HDM2-inhibitor complexes and uses thereof

ABSTRACT

The present invention includes crystallized HDM2 peptides as well as descriptions of the X-ray diffraction patterns of the crystals. The diffraction patterns allow the three dimensional structure of HDM2 to be determined at atomic resolution so that ligand binding sites on HDM2 can be identified and the interactions of ligands with HDM2 amino acid residues can be modeled. Models prepared using such maps permit the design of ligands which can function as active agents which include, but are not limited to, those that function as inhibitors of MDM2 and HDM2 oncoproteins.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to Application No. 60/418,350filed on Oct. 16, 2002.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH DEVELOPMENT

[0002] Not applicable.

FIELD OF THE INVENTION

[0003] The present invention generally pertains to the fields ofmolecular biology, protein crystallization, X-ray diffraction analysis,three-dimensional structural determination, molecular modeling andstructure based rational drug design. The present invention providescrystallized HDM2 peptides as well as descriptions of the X-raydiffraction patterns. The X-ray diffraction patterns of the crystals inquestion are of sufficient resolution so that the three-dimensionalstructure of HDM2 can be determined at atomic resolution, ligand bindingsites on HDM2 can be identified, and the interactions of ligands withHDM2 amino acid residues can be modeled.

[0004] The high resolution maps provided by the present invention andthe models prepared using such maps also permit the design of ligandswhich can function as active agents. Thus, the present invention hasapplications to the design of active agents which include, but are notlimited to, those that find use as inhibitors of MDM2 and HDM2oncoproteins.

BACKGROUND OF THE INVENTION

[0005] HDM2: Structure and Function

[0006] HDM2 (human double minute 2 protein) is the expression product ofhdm2, an oncogene that is overexpressed in a subset of human tumorsincluding soft tissue sarcomas, glioblastomas and mammary carcinomas(Oliner, J. D. et al., Nature, 358(6381):80-83 (1992); Reifenberger, G.et al., Cancer Res., 53:2736-2739 (1993); Bueso-Ramos, C. E. et al.,Breast Canc. Res. Treat., 37(2):179-188 (1996)).

[0007] Functional characterization of this oncogene revealed aninteraction between HDM2 and p53, a tumor suppressor central to cellgrowth arrest and apoptosis (Momand, G. P. et al., Cell, 69:1237(1992)). HDM2 is a transcriptional target of p53, and as such, HDM2 andp53 form a precisely regulated loop (Wu, X. et al., Genes and Dev.,7:1126-1132 (1992)). HDM2 is further regulated by ubiquitination and bycomplex formation with Arf, which sequesters HDM2 to the nucleolus (Tao,W. and Levine, A. J., Proc. Natl. Acad. Sci. USA, 96(12):6937-6941(1999); Weber, J. D. et al., Nat. Cell Biol., 1(1):20-26 (1999)).

[0008] There are several lines of evidence that suggest that HDM2 canalso function independently of p53. Splice variants of HDM2 notcontaining the p53-binding domain have been found in human tumors andhave been shown to possess transforming ability (Sigalas, I. et al. Nat.Med. 2(8):912-917 (1996)). In vivo studies have also demonstrated thatthe spectrum of tumors that develop in transgenic mice overexpressingHDM2 is different from the spectrum found in p53-null mice and that HDM2can drive sarcomagenesis in p53-null animals (Jones, S. N. et al., Proc.Natl. Acad. Sci. USA, 95(26):15608-15612 (1998)). Lastly, other bindingpartners of HDM2 could assist HDM2 function along an oncogenic pathway,for example, HDM2 inhibition of MTBP-induced p53-independent G1 arrest(Boyd, M. T. et al., J. Biol. Chem. 275(41):31883-31890 (2000)).

[0009] Reports of enhanced tumor cell death following hdm2 inhibition byantisense nucleotides (Chen, L. et al., Proc. Natl. Acad Sci. USA,95(1):195-200 (1998); Chen, L. et al., Mol. Med., 5(1):21-34 (1999);Tortora, G. et al., Int. J. Cancer, 88(5):804-809 (2000)) andHDM2-binding mini-proteins (Bottger, A. et al., Curr. Biol.,7(11):860-869 (1997)) substantiate a prediction that inhibition of HDM2will activate p53 and in turn trigger apoptosis. Following on this idea,a small molecule inhibitor generated against the p53 binding groove ofHDM2 would be expected to prevent the interaction of the two proteinsand induce p53 activity. It has been further suggested that inhibitingthe interaction between p53 and HDM2 will act additively orsynergistically with standard chemotherapeutic agents in the treatmentof neoplasm, and this too is supported by work utilizing antisense hdm2constructs (Wang, H. et al., Clin. Canc. Res. 7(11):3613-3624 (2001)).

[0010] MDM2: Structure and Function

[0011] mdm2, the murine homolog of HDM2 was originally found on mousedouble minute chromosomes and was initially identified as one of threegenes amplified in a tumorigenic cell line (Cahilly-Snyder., L. et al.,Somatic Cell Mol. Genet. 13:235-244 (1987)). Its protein product wassubsequently found to form a complex with p53, which was first observedin a rat fibroblast cell line (Clone 6) previously transfected with atemperature sensitive mouse p53 gene (Michalovitz, D. et al., Cell62:671-680 (1990)). The rat cell line grew well at 37° C. but exhibiteda G1 arrest when shifted down to 32° C., which was entirely consistentwith an observed temperature dependent switch in p53 conformation andactivity. However, the p53-MDM2 complex was only observed in abundanceat 32° C., at which temperature p53 was predominantly in a functional or“wild-type” form (Barak, Y. et al., EMBO J. 11:2115-2121 (1992) andMomand, J. et al., Cell 69:1237-1245 (1992)). By shifting the rat cellline down to 32° C. and blocking de novo protein synthesis it was shownthat only “wild-type” p53 induced expression of the mdm2 gene, therebyaccounting for the differential abundance of the complex in terms of p53transcriptional activity (Barak, Y. et al., EMBO J. 12:461-468 (1993)).The explanation was further developed by the identification of a DNAbinding site for wild-type p53 within the first intron of the mdm2 gene(Wu, X. et al., Genes Dev. 7:1126-1132 (1993)). Reporter constructsemploying this p53 DNA binding site revealed that they were inactivatedwhen wild-type p53 was co-expressed with MDM2.

[0012] This inhibition of the transcriptional activity of p53 may becaused by MDM2 blocking the activation domain of p53 and/or the DNAbinding site. Consequently, it was proposed that mdm2 expression isautoregulated, via the inhibitory effect of MDM2 protein on thetranscriptional activity of wild-type p53. This p53-mdm2 autoregulatoryfeedback loop provided a novel insight as to how cell growth might beregulated by p53. Up to a third of human sarcomas are considered toovercome p53-regulated growth control by amplification of the mdm2 gene(Oliner, J. D. et al., Nature 358:80-83 (1992)). Hence, the interactionbetween p53 and MDM2 represents a key potential therapeutic target.

[0013] p53: Interaction with HDM2 and MDM2

[0014] p53 is a transcription factor for a number of proteins that causecell cycle arrest or cell death by apoptosis, such as p21, 14-3-3σ, andbax. The level and transcriptional activity of p53 are increased bydamage to cellular DNA. The MDM2 protein inhibits p53 function bybinding to an amphipathic N-terminal helix of p53, abrogating theinteraction of p53 with other proteins and its trans activationactivity. The interaction with MDM2 also targets p53 for ubiquitindependent protein degradation. MDM2 exhibits p53 independent effects oncell cycling as well, possibly by direct interaction with some of thedownstream effectors such as pRB and EF2 (Reviewed in Zhang, R. andWang, H., Cur. Pharm. Des. 6:393-416 (2000)).

[0015] Mutations of the p53 protein occur in 50% of all human cancers(reviewed in Agarwal, M. L. et al. J. Biol. Chem. 273:1-4 (1998);Levine, A. J., Cell 88:323-331(1997); and, references cited in Oren, M.,J. Biol. Chem. 274:36031-36034 (1999)). Under normal circumstances, p53is latent and a very labile protein, which turns over with a very shorthalf-life of a few minutes (Rogel, A. et al., Mol. Cell. Biol.5:2851-2855 (1985)). DNA damage or stress induces a remarkable increasein the stability of p53 (Kastan, M. B. et al., Cancer Res. 51:6304-6311(1991)). Furthermore, these signals, also activate the function of p53as a transcriptional activator of the apoptotic machinery, a functionnormally suppressed by autoregulatory inhibition of its transactivationdomain. The amount of p53 present in the cell is tightly regulated by anegative feedback loop between p53 and the oncogene hdm2.

[0016] p53 is located in the cell nucleus and induces the expression ofhdm2 through its transactivation domain. Expressed hdm2 subsequentlybinds to residues 19-26 of the p53 transactivation domain, inactivatesit (Chen, J. et al., Mol. Cell. Biol. 16:2445-2452 (1996); Haupt, Y. etal., EMBO J. 15:1596-1606 (1996); Momand, J. et al., Cell 69:1237-1245(1992)) and blocks recruitment of transcription factors necessary forgene expression (Lu, H. et al., PNAS 92:5154-5158 (1995); Thut, C. J. etal., Science 267:00-104 (1995)). Furthermore, the p53-hdm2-complex isshuttled to the cytoplasm where degradation occurs. This tight controlthrough negative feedback is critical for the survival of the organism.Inactivation of hdm2 in hdm2-knockout mice leads to early embryonallethality, but is completely prevented by simultaneous inactivation ofp53 (Jones, S. N. et al., Nature 378:206-208 (1995); Montes de Oca Luna,R. et al., Nature 378:203-206 (1995)). On the other hand, excessiveexpression of hdm2 can lead to constitutive inhibition of p53 andpromote cancer. Excess HDM2 also promotes cancer independently of p53(Lundgren, K. et al., Genes and Dev. 11:714-725 (1997); Sun, P. et al.,Science 282:2270-2272 (1998)).

[0017] As discussed above, inhibition of the interaction between HDM2and p53 is an attractive target for cancer therapy (Lane, D. P., TIBS22: 372-374 (1997)). It has been shown that inhibition of the complexformation between p53 and HDM2 raises the levels of p53 in the cell(Bottger, A. et al., Current Biol. 7:860-869 (1997)). Also, blockingHDM2 from binding p53 would be therapeutically useful in restoring cellcycle control to cells that overexpress HDM2 as a front line cancertreatment. More generally, inhibition of HDM2 may increase theeffectiveness of chemotherapy and radiation in p53 normal cancers byenhancing apoptosis and growth arrest signaling pathways. This approachmay render tumor cells containing functional p53 more susceptible tochemotherapeutic agents.

[0018] One method of identifying inhibitors of the p53/HDM2 proteincomplex is to determine the amino acid specificities of HDM2 bindingpockets by crystallography in order to establish a model for theinteraction. Using this method, Kussie et al. identified p53 basedpeptide antagonists (Kussie, P. H. et al., Science 274:948-953 (1996)).A crystal structure of a truncated form of HDM2 (residues 17-125) and a15′mer peptide derived from the N-terminal transactivation domain of p53was published by Kussie et al. (Kussie et al., (1996)). Kussie et al.also published a crystal structure of MDM2 (Kussie et al., (1996))derived from Xenopus laevis (residues 13 to 118), having a 71% sequenceidentity towards HDM2. Based on molecular modeling, Garcia-Echeverrìa etal. published a model of an 8′mer peptidomimetic, derived from the15′mer wild-type p53 peptide, bound to the N-terminal domain of hdm2(Garcia-Echeverrìa, C. et al., J. Med. Chem. 43:3205-3208 (2000)). Nocrystal structure of HDM2 with inhibitory compounds, such as smallmolecule inhibitors, for example, or other peptides is believed to havebeen disclosed.

[0019] Therefore, a need continues to exist for the development ofmodeling systems to design and select potent, small molecules thatinhibit the interactions between HDM2 (and homologs thereof) and naturalbinding ligands such as p53.

SUMMARY OF THE INVENTION

[0020] The present invention includes methods of producing and usingthree-dimensional structure information derived from human MDM2 protein(HDM2) and inhibitory compounds which form a complex with HDM2 andprevent HDM2 from interacting with the p53 protein. The presentinvention also includes specific crystallization conditions to obtaincrystals of the inhibitor-HDM2 complex. The crystals are subsequentlyused to obtain a 3-dimensional structure of the complex using X-raycrystallography (or NMR) and the obtained data is used for rational drugdiscovery design with the aim to improve the complex formation betweenHDM2 and the inhibitor, and, also to improve the inhibition of thebinding of HDM2 to p53.

[0021] The present invention includes a crystal comprising HDM2, or afragment, or target structural motif or derivative thereof, and aligand, wherein the ligand is a small molecule inhibitor. In anotherembodiment, the crystal has a spacegroup selected from the groupconsisting of a trigonal spacegroup of P3₂21 and a tetragonal spacegroupP4₃2₁2. The present invention also includes a crystal comprising HDM2which comprises a peptide having at least 95% sequence identity to SEQID NO. 2.

[0022] In another aspect of the invention, the invention includes acomputer system comprising: (a) a database containing information on thethree dimensional structure of a crystal comprising HDM2, or a fragmentor a target structural motif or derivative thereof, and a ligand,wherein the ligand is a small molecule inhibitor, stored on a computerreadable storage medium; and, (b) a user interface to view theinformation.

[0023] The present invention also includes a method of evaluating thepotential of an agent to associate with HDM2 comprising: (a) exposingHDM2 to the agent; and (b) detecting the association of said agent toHDM2 amino acid residues Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷thereby evaluatingthe potential.

[0024] The invention further includes a method of evaluating thepotential of an agent to associate with the peptide having aa¹⁶-SEQ IDNO: 2, comprising: (a) exposing aa¹⁶-SEQ ID NO: 2 to the agent; and (b)detecting the level of association of the agent to aa ¹⁶-SEQ ID NO: 2,thereby evaluating the potential.

[0025] Further included in the present invention is a method ofidentifying a potential agonist or antagonist against HDM2 comprising:(a) employing the three dimensional structure of HDM2 cocrystallizedwith a small molecule inhibitor to design or select said potentialagonist or antagonist.

[0026] The instant invention comprises a method of locating theattachment site of an inhibitor to HDM2, comprising: (a) obtaining X-raydiffraction data for a crystal of HDM2; (b) obtaining X-ray diffractiondata for a complex of HDM2 and an inhibitor; (c) subtracting the X-raydiffraction data obtained in step (a) from the X-ray diffraction dataobtained in step (b) to obtain the difference in the X-ray diffractiondata; (d) obtaining phases that correspond to X-ray diffraction dataobtained in step (a); (e) utilizing the phases obtained in step (d) andthe difference in the X-ray diffraction data obtained in step (c) tocompute a difference Fourier image of the inhibitor; and, (f) locatingthe attachment site of the inhibitor to HDM2 based on the computationsobtained in step (e).

[0027] The present invention further comprises a method of obtaining amodified inhibitor comprising: (a) obtaining a crystal comprising HDM2and an inhibitor; (b) obtaining the atomic coordinates of the crystal;(c) using the atomic coordinates and one or more molecular modelingtechniques to determine how to modify the interaction of the inhibitorwith HDM2; and, (d) modifying the inhibitor based on the determinationsobtained in step (c) to produce a modified inhibitor.

[0028] In another aspect of the invention, the invention includes anisolated protein fragment comprising a binding pocket or active sitedefined by structure coordinates of HDM2 amino acid residues Ser¹⁷,Ile¹⁹, Leu⁸² and Arg⁹⁷.

[0029] In another aspect of the invention, the invention includes anisolated nucleic acid molecule encoding the fragment which comprises abinding pocket or active site defined by structure coordinates of HDM2amino acid residues Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷. In another aspect ofthe invention, the invention includes a method of screening for an agentthat associates with HDM2, comprising: (a) exposing a protein moleculefragment to the agent; and (b) detecting the level of association of theagent to the fragment. In another aspect of the invention, the inventionincludes a kit comprising a protein molecule fragment.

[0030] The invention additionally comprises a method for the productionof a crystal complex comprising an HDM2 polypeptide-ligand comprising:(a) contacting the HDM2 polypeptide with said ligand in a suitablesolution comprising PEG and NaSCN; and, b) crystallizing said resultingcomplex of HDM2 polypeptide-ligand from said solution.

[0031] The invention further includes a method for the production of acrystal comprising HDM2 and a ligand wherein the ligand is a smallmolecule inhibitor comprising crystallizing a peptide comprising asequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO: 3 and SEQ ID NO:4 with a potential inhibitor.

[0032] The instant invention includes a method for identifying apotential inhibitor of HDM2 comprising: a) using a three dimensionalstructure of HDM2 as defined by atomic coordinates according to table 1or table 2; b) replacing one or more HDM2 amino acids selected fromSer¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷ in said three-dimensional structure with adifferent amino acid to produce a modified HDM2; c) using saidthree-dimensional structure to design or select said potentialinhibitor; d) synthesizing said potential inhibitor; and, e) contactingsaid potential inhibitor with said modified HDM2 in the presence of asubstrate to test the ability of said potential inhibitor to inhibitHDM2 or said modified HDM2. Also included in the invention is aninhibitor identified by the method.

BRIEF DESCRIPTION OF THE DRAWINGS

[0033]FIG. 1 Ribbon representation of HDM2 bound to compound 338437.

[0034]FIG. 2 Fit of compound 338437 into the active site of HDM2represented as a molecular surface.

[0035]FIG. 3 Ribbon representation of a superposition between hdm2 inthe trigonal crystal form and in the tetragonal form. The RMS deviationbetween C-alpha atom positions is 0.25 Angstroms.

DETAILED DESCRIPTION OF THE INVENTION

[0036] Definitions

[0037] As is generally the case in biotechnology and chemistry, thedescription of the present invention has required the use of a number ofterms of art. Although it is not practical to do so exhaustively,definitions for some of these terms are provided here for ease ofreference. Unless defined otherwise, all technical and scientific termsused herein have the same meaning as commonly understood by one ofordinary skill in the art to which this invention belongs. Definitionsfor other terms also appear elsewhere herein. However, the definitionsprovided here and elsewhere herein should always be considered indetermining the intended scope and meaning of the defined terms.Although any methods and materials similar or equivalent to thosedescribed herein can be used in the practice of the present invention,the preferred methods and materials are described.

[0038] As used herein, the term “atomic coordinates” or “structurecoordinates” refers to mathematical coordinates that describe thepositions of atoms in crystals of HDM2 in Protein Data Bank (PDB)format, including X, Y, Z and B, for each atom. The diffraction dataobtained from the crystals are used to calculate an electron density mapof the repeating unit of the crystal. The electron density maps may beused to establish the positions (i.e. coordinates X, Y and Z) of theindividual atoms within the crystal. Those of skill in the artunderstand that a set of structure coordinates determined by X-raycrystallography is not without standard error. For the purpose of thisinvention, any set of structure coordinates for HDM2 from any sourcehaving a root mean square deviation of non-hydrogen atoms of less thanabout 1.5 Å when superimposed on the non-hydrogen atom positions of thecorresponding atomic coordinates of Table 1 or Table 2 are consideredsubstantially identical or homologous. In a more preferred embodiment,any set of structure coordinates for HDM2 from any source having a rootmean square deviation of non-hydrogen atoms of less than about 0.75 Åwhen superimposed on the non-hydrogen atom positions of thecorresponding atomic coordinates of Table 1 or Table 2 are consideredsubstantially identical or homologous.

[0039] The term “atom type” refers to the chemical element whosecoordinates are measured. The first letter in a column in Table 1identifies the element.

[0040] The terms “X,” “Y” and “Z” refer to thecrystallographically-defined atomic position of the element measuredwith respect to the chosen crystallographic origin. The term “B” refersto a thermal factor that measures the mean variation of an atom'sposition with respect to its average position.

[0041] As used herein, the term “crystal” refers to anythree-dimensional ordered array of molecules that diffracts X-rays.

[0042] As used herein, the term “carrier” in a composition refers to adiluent, adjuvant, excipient, or vehicle with which the product ismixed.

[0043] As used herein, the term “composition” refers to the combining ofdistinct elements or ingredients to form a whole. A compositioncomprises more than one element or ingredient. For the purposes of thisinvention, a composition will often, but not always, comprise a carrier.

[0044] As used herein, “mdm2” is used to mean the murine double minute 2gene, and homologous genes found in other animals.

[0045] As used herein, “MDM2” is used to mean a protein obtained as aresult of expression of the mdm2 oncogene. Within the meaning of thisterm, it will be understood that MDM2 encompasses all proteins encodedby mdm2, mutants thereof, conservative amino acid substitutions,alternative splice proteins thereof, and phosphorylated proteinsthereof. Additionally, as used herein, it will be understood that theterm “MDM2” includes MDM2 homologues of other animals.

[0046] As used herein, “hdm2” is used to mean the human gene, which ishomologous to the mouse mdm2 gene.

[0047] As used herein, “HDM2” is used to mean a protein obtained as aresult of expression of the hdm2 oncogene. Within the meaning of thisterm, it will be understood that HDM2 encompasses all proteins encodedby hdm2, mutants thereof, conservative amino acid substitutions,alternative splice proteins thereof, and phosphorylated proteinsthereof. As an example, HDM2 includes the protein comprising SEQ ID NO:2 and variants thereof comprising at least about 70% amino acid sequenceidentity to SEQ ID NO: 2, or preferably 80%, 85%, 90% and 95% sequenceidentity to SEQ ID NO: 2, or more preferably, at least about 95% or moresequence identity to SEQ ID NO: 2.

[0048] As used herein, the term “SAR,” an abbreviation forStructure-Activity Relationships, collectively refers to thestructure-activity/structure property relationships pertaining to therelationship(s) between a compound's activity/properties and itschemical structure.

[0049] As used herein, the term “molecular structure” refers to thethree dimensional arrangement of molecules of a particular compound orcomplex of molecules (e.g., the three dimensional structure of HDM2 andligands that interact with HDM2).

[0050] As used herein, the term “molecular modeling” refers to the useof computational methods, preferably computer assisted methods, to drawrealistic models of what molecules look like and to make predictionsabout structure activity relationships of ligands. The methods used inmolecular modeling range from molecular graphics to computationalchemistry.

[0051] As used herein, the term “molecular model” refers to the threedimensional arrangement of the atoms of a molecule connected by covalentbonds or the three dimensional arrangement of the atoms of a complexcomprising more than one molecule, e.g., a protein-ligand complex.

[0052] As used herein, the term “molecular graphics” refers to 3Drepresentations of the molecules, for instance, a 3D representationproduced using computer assisted computational methods.

[0053] As used herein, the term “computational chemistry” refers tocalculations of the physical and chemical properties of the molecules.

[0054] As used herein, the term “molecular replacement” refers to amethod that involves generating a preliminary model of a crystal of HDM2whose coordinates are unknown, by orienting and positioning the saidatomic coordinates described in the present invention so as best toaccount for the observed diffraction pattern of the unknown crystal.Phases can then be calculated from this model and combined with theobserved amplitudes to give an approximate Fourier synthesis of thestructure whose coordinates are unknown. (Rossmann, M. G., ed., “TheMolecular Replacement Method”, Gordon & Breach, New York, 1972).

[0055] As used herein, the term “homolog” refers to the HDM2 proteinmolecule or the nucleic acid molecule which encodes the protein, or afunctional domain from said protein from a first source having at leastabout 30%, 40% or 50% sequence identity, or at least about 60%, 70% or75% sequence identity, or at least about 80% sequence identity, or morepreferably at least about 85% sequence identity, or even more preferablyat least about 90% sequence identity, and most preferably at least about95%, 97% or 99% amino acid or nucleotide sequence identity, with theprotein, encoding nucleic acid molecule or any functional domainthereof, from a second source. The second source may be a version of themolecule from the first source that has been genetically altered by anyavailable means to change the primary amino acid or nucleotide sequenceor may be from the same or a different species than that of the firstsource.

[0056] As used herein, the term “active site” refers to regions on HDM2or a structural motif of HDM2 that are directly involved in the functionor activity of HDM2.

[0057] As used herein, the terms “binding site” or “binding pocket”refer to a region of HDM2 or a molecular complex comprising HDM2 that,as a result of the primary amino acid sequence of HDM2 and/or itsthree-dimensional shape, favorably associates with another chemicalentity or compound including ligands or inhibitors.

[0058] For the purpose of this invention, any active site, binding siteor binding pocket defined by a set of structure coordinates for HDM2 orfor a homolog of HDM2 from any source having a root mean squaredeviation of non-hydrogen atoms of less than about 1.5 Å whensuperimposed on the non-hydrogen atom positions of the correspondingatomic coordinates of Table 1 or Table 2 are considered substantiallyidentical or homologous. In a more preferred embodiment, any set ofstructure coordinates for HDM2 or a homolog of HDM2 from any sourcehaving a root mean square deviation of non-hydrogen atoms of less thanabout 0.75 Å when superimposed on the non-hydrogen atom positions of thecorresponding atomic coordinates of Table 1 or Table 2 are consideredsubstantially identical or homologous.

[0059] The tem “root mean square deviation” means the square root of thearithmetic mean of the squares of the deviations from the mean.

[0060] As used herein, the term “amino acids” refers to the L-isomers ofthe naturally occuring amino acids. The naturally occurring amino acidsare glycine, alanine, valine, leucine, isoleucine, serine, methionine,threonine, phenylalanine, tyrosine, tryptophan, cysteine, proline,histidine, aspartic acid, asparagine, glutamic acid, glutamine,γ-carboxylglutamic acid, arginine, ornithine, and lysine. Unlessspecifically indicated, all amino acids are referred to in thisapplication are in the L-form.

[0061] As used herein, the term “nonnatural amino acids” refers to aminoacids that are not naturally found in proteins. For example,selenomethionine.

[0062] As used herein, the term “positively charged amino acid” includesany amino acids having a positively charged side chain under normalphysiological conditions. Examples of positively charged naturallyoccurring amino acids are arginine, lysine, and histidine.

[0063] As used herein, the term “negatively charged amino acid” includesany amino acids having a negatively charged side chains under normalphysiological conditions. Examples of negatively charged naturallyoccurring amino acids are aspartic acid and glutamic acid.

[0064] As used herein, the term “hydrophobic amino acid” includes anyamino acids having an uncharged, nonpolar side chain that is relativelyinsoluble in water. Examples of naturally occurring hydrophobic aminoacids are alanine, leucine, isoleucine, valine, proline, phenylalanine,tryptophan, and methionine.

[0065] As used herein, the term “hydrophilic amino acid” refers to anyamino acids having an uncharged, polar side chain that is relativelysoluble in water. Examples of naturally occurring hydrophilic aminoacids are serine, threonine, tyrosine, asparagine, glutamine andcysteine.

[0066] As used herein, the term “hydrogen bond” refers to twohydrophilic atoms (either O or N), which share a hydrogen that iscovalently bonded to only one atom, while interacting with the other.

[0067] As used herein, the term “hydrophobic interaction” refers tointeractions made by two hydrophobic residues or atoms (such as C).

[0068] As used herein, the term “conjugated system” refers to more thantwo double bonds are adjacent to each other, in which electrons arecompletely delocalized with the entire system. This also includes andaromatic residues.

[0069] As used herein, the term “aromatic residue” refers to amino acidswith side chains having a delocalized conjugated system. Examples ofaromatic residues are phenylalanine, tryptophan, and tyrosine.

[0070] As used herein, the phrase “inhibiting the binding” refers topreventing or reducing the direct or indirect association of one or moremolecules, peptides, proteins, enzymes, or receptors, or preventing orreducing the normal activity of one or more molecules, peptides,proteins, enzymes or receptors, e.g., preventing or reducing the director indirect association of HDM2 and p53.

[0071] As used herein, the term “competitive inhibitor” refers toinhibitors that bind to HDM2 at the same sites as its binding partner(s)(e.g., p53), thus directly competing with them. Competitive inhibitionmay, in some instances, be reversed completely by increasing thesubstrate concentration.

[0072] As used herein, the term “uncompetitive inhibitor” refers to onethat inhibits the functional activity of HDM2 by binding to a differentsite than does its substrate(s) e.g. (p53).

[0073] As used herein, the term “non-competitive inhibitor” refers toone that can bind to either the free or p53 bound form of HDM2.

[0074] Those of skill in the art may identify inhibitors as competitive,uncompetitive, or non-competitive by computer fitting enzyme kineticdata using standard methods. See, for example, Segel, I. H., EnzymeKinetics, J. Willey & Sons, (1975).

[0075] As used herein, the term “R or S-isomer” refers to two possiblestereroisomers of a chiral carbon according to the Cahn-Ingold-Prelogsystem adopted by International Union of Pure and Applied Chemistry(IUPAC). Each group attached to the chiral carbon is first assigned to apreference or priority a, b, c, or d on the basis of the atomic numberof the atom that is directly attached to the chiral carbon. The groupwith the highest atomic number is given the highest preference a, thegroup with next highest atomic number is given the next highestpreference b; and so on. The group with the lowest preference (d) isthen directed away from the viewer. If the trace of a path from a to bto c is counter clockwise, the isomer is designated (S); in the oppositedirection, clockwise, the isomer is designated (R).

[0076] As used herein, the term “ligand” refers to any molecule, orchemical entity which binds with or to HDM2, a subunit of HDM2, a domainof HDM2, a target structual motif of HDM2 or a fragment of HDM2. Thus,ligands include, but are not limited to, small molecule inhibitors, forexample.

[0077] As used herein, the term “small molecule inhibitor” refers tocompounds useful in the present invention having measurable MDM2 or HDM2inhibiting activity. In addition to small organic molecules, peptides,antibodies, cyclic peptides and peptidomimetics are contemplated asbeing useful in the disclosed methods. Excluded from the invention arethe p53 peptides disclosed in Kussie et al., Garcia-Echeverrìa et al.,and the peptides derived from phage display which inhibit the binding ofmdm2 to p53 (Böttger, V. A., et al., Oncogene 13(10): 2141-2147 (1996)).Preferred inhibitors are small molecules, preferably less than 700Daltons, and more preferably less than 450 Daltons. Examples of classesof compounds having this property include compounds disclosed in U.S.Provisional Application No. 60/275,629; in U.S. Provisional ApplicationNo. 60/331,235; in U.S. Provisional Application No. 60/379,617; and, inU.S. application Ser. No. 10/097,249, incorporated herein in theirentirety.

[0078] As used herein the terms “bind,” “binding,” “bond,” or “bonded”when used in reference to the association of atoms, molecules, orchemical groups, refer to any physical contact or association of two ormore atoms, molecules, or chemical groups.

[0079] As used herein, the terms “covalent bond” or “valence bond” referto a chemical bond between two atoms in a molecule created by thesharing of electrons, usually in pairs, by the bonded atoms.

[0080] As used herein, “noncovalent bond” refers to an interactionbetween atoms and/or molecules that does not involve the formation of acovalent bond between them.

[0081] As used herein, the term “native protein” refers to a proteincomprising an amino acid sequence identical to that of a proteinisolated from its natural source or organism.

SPECIFIC EMBODIMENTS DETAILED EMBODIMENTS

[0082] The present invention includes a crystal comprising HDM2, or afragment, or target structural motif or derivative thereof, and aligand, wherein the ligand is a small molecule inhibitor. In oneembodiment, the fragment or derivative thereof is a peptide selectedfrom the group consisting of SEQ ID NO: 1 (amino acid sequence of fulllength HDM2), SEQ ID NO: 2 (amino acid residues 17-111 of SEQ ID NO: 1),SEQ ID NO. 3 (amino acid residues 23-114 of SEQ ID NO: 1) and SEQ ID NO.4 (Gly¹⁶-SEQ ID NO: 2).

[0083] In another embodiment, the crystal has a spacegroup selected fromthe group consisting of a trigonal spacegroup of P3₂21 and a tetragonalspacegroup of P4₃2₁2. In a different embodiment, the crystal effectivelydiffracts X-rays for determination of atomic coordinates to a resolutionof at least about 3.0 Å. In a preferred embodiment, the ligand is incrystalline form. In a highly preferred embodiment, the ligand isselected from the group consisting of(4-Chloro-phenyl)-[3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-aceticacid;[8-Chloro-3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-(4-chloro-phenyl)-aceticacid, and, derivatives thereof.

[0084] The present invention also includes a crystal comprising HDM2which comprises a peptide having at least 95% sequence identity to SEQID NO. 2. In a preferred embodiment, the crystal comprising SEQ ID NO: 2comprises an atomic structure characterized by the coordinates of Table1 or Table 2. In another preferred embodiment, the crystal comprises aunit cell selected from the group consisting of: a cell havingdimensions of about 98.6 Å, 98.6 Å and 74.7 Å, and about alpha=90°,beta=90° and gamma=120°; and, a cell having dimensions of about 54.3 Å,54.3 Å, 83.3 Å and about alpha=90°, beta=90° and gamma=90°.

[0085] In another aspect of the invention, the invention includes acomputer system comprising: (a) a database containing information on thethree dimensional structure of a crystal comprising HDM2, or a fragmentor a target structural motif or derivative thereof, and a ligand,wherein the ligand is a small molecule inhibitor, stored on a computerreadable storage medium; and, (b) a user interface to view theinformation. In one embodiment, the information comprises diffractiondata obtained from a crystal comprising SEQ ID NO:2. In anotherembodiment, the information comprises an electron density map of acrystal form comprising SEQ ID NO:2. In a different embodiment, theinformation comprises the structure coordinates of Table 1 or Table 2 orhomologous structure coordinates comprising a root mean square deviationof non-hydrogen atoms of less than about 1.5 A when superimposed on thenon-hydrogen atom positions of the corresponding atomic coordinates ofTable 1 or Table 2. In a preferred embodiment, the information comprisesstructure coordinates comprising a root mean square deviation ofnon-hydrogen atoms of less than about 0.75 Å when superimposed on thenon-hydrogen atom positions of the corresponding atomic coordinates ofTable 1 or Table 2. In a highly preferred embodiment, the informationcomprises the structure coordinates for amino acids Ser¹⁷, Ile¹⁹ , Leu⁸²and Arg⁹⁷ according to Table 1 or Table 2 or similar structurecoordinates for said amino acids comprising a root mean square deviationof non-hydrogen atoms of less than about 1.5 Å when superimposed on thenon-hydrogen atom positions of the corresponding atomic coordinates ofTable 1 or Table 2. In another embodiment, the information furthercomprises the structure coordinates for amino acids Val⁵³, Leu⁵⁴, Phe⁵⁵,Leu⁵⁷, Gly⁵⁸, Gln⁵⁹, Ile⁶¹, Met⁶², Tyr⁶⁷, Gln⁷², His⁷³, Ile⁷⁴, Val⁷⁵,Phe⁸⁶, Phe⁹¹, Val⁹³, Lys⁹⁴, Glu⁹⁵, His⁹⁶, Ile⁹⁹, Tyr¹⁰⁰, Ile¹⁰³according to Table 1 or Table 2 or similar structure coordinates forsaid amino acids comprising a root mean square deviation of non-hydrogenatoms of less than about 1.5 Å when superimposed on the non-hydrogenatom positions of the corresponding atomic coordinates of Table 1 orTable 2.

[0086] The present invention also includes a method of evaluating thepotential of an agent to associate with HDM2 comprising: (a) exposingHDM2 to the agent; and (b) detecting the association of said agent toHDM2 amino acid residues Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷ therebyevaluating the potential. In one embodiment of the invention, the agentis a virtual compound. In another embodiment of the invention, step (a)comprises comparing the atomic structure of the compound to the threedimensional structure of HDM2. In a different embodiment, the comparingcomprises employing a computational means to perform a fitting operationbetween the compound and at least one binding site of HDM2. In apreferred embodiment, the binding site is defined by structurecoordinates for amino acids Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷ according toTable 1 or Table 2 similar structure coordinates for said amino acidscomprising a root mean square deviation of non-hydrogen atoms of lessthan about 1.5 Å when superimposed on the non-hydrogen atom positions ofthe corresponding atomic coordinates of Table 1 or Table 2. In anotherdifferent embodiment, the binding site is further defined by structurecoordinates for amino acids Val⁵³, Leu⁵⁴, Phe⁵⁵, Leu⁵⁷, Gly⁵⁸, Gln⁵⁹,Ile⁶¹, Met⁶², Tyr⁶⁷, Gln⁷², His⁷³, Ile⁷⁴, Val⁷⁵, Phe⁸⁶, Phe⁹¹, Val⁹³,Lys⁹⁴, Glu⁹⁵, His⁹⁶, Ile⁹⁹, Tyr¹⁰⁰, Ile¹⁰³ according to Table 1 or Table2 or similar structure coordinates for said amino acids comprising aroot mean square deviation of non-hydrogen atoms of less than about 1.5Å when superimposed on the non-hydrogen atom positions of thecorresponding atomic coordinates of Table 1 or Table 2. In a highlypreferred embodiment, the agent is exposed to crystalline SEQ ID NO:2and the detecting of step (b) comprises determining the threedimensional structure of the agent-SEQ ID NO: 2 complex.

[0087] The invention further includes a method of evaluating thepotential of an agent to associate with the peptide having aa¹⁶-SEQ IDNO: 2, comprising: (a) exposing aa¹⁶-SEQ ID NO: 2 to the agent; and (b)detecting the level of association of the agent to aa ¹⁶-SEQ ID NO: 2,thereby evaluating the potential. In one embodiment, the agent is avirtual compound.

[0088] The present invention includes a method of identifying apotential agonist or antagonist against HDM2 comprising: (a) employingthe three dimensional structure of HDM2 cocrystallized with a smallmolecule inhibitor to design or select said potential agonist orantagonist. In one embodiment, the three dimensional structurecorresponds to the atomic structure characterized by the coordinates ofTable 1 or similar structure coordinates comprising a root mean squaredeviation of non-hydrogen atoms of less than about 1.5 Å whensuperimposed on the non-hydrogen atom positions of the correspondingatomic coordinates of Table 1 or Table 2. In a different embodiment, themethod further comprises the steps of: (b) synthesizing the potentialagonist or antagonist; and (c) contacting the potential agonist orantagonist with HDM2.

[0089] The instant invention comprises a method of locating theattachment site of an inhibitor to HDM2, comprising: (a) obtaining X-raydiffraction data for a crystal of HDM2; (b) obtaining X-ray diffractiondata for a complex of HDM2 and an inhibitor; (c) subtracting the X-raydiffraction data obtained in step (a) from the X-ray diffraction dataobtained in step (b) to obtain the difference in the X-ray diffractiondata; (d) obtaining phases that correspond to X-ray diffraction dataobtained in step (a); (e) utilizing the phases obtained in step (d) andthe difference in the X-ray diffraction data obtained in step (c) tocompute a difference Fourier image of the inhibitor; and, (f) locatingthe attachment site of the inhibitor to HDM2 based on the computationsobtained in step (e).

[0090] The present invention further comprises a method of obtaining amodified inhibitor comprising: (a) obtaining a crystal comprising HDM2and an inhibitor; (b) obtaining the atomic coordinates of the crystal;(c) using the atomic coordinates and one or more molecular modelingtechniques to determine how to modify the interaction of the inhibitorwith HDM2; and, (d) modifying the inhibitor based on the determinationsobtained in step (c) to produce a modified inhibitor. In one embodiment,the crystal comprises a peptide selected from the group consisting of: apeptide having SEQ ID NO: 2; a peptide having SEQ ID NO: 3 and a peptidehaving SEQ ID NO:4. In a different embodiment, the one or more molecularmodeling techniques are selected from the group consisting of graphicmolecular modeling and computational chemistry. In a preferredembodiment, step (a) comprises detecting the interaction of theinhibitor to HDM2 amino acid residues Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷. Inanother embodiment of the invention, the invention includes an HDM2inhibitor identified by this method.

[0091] In another aspect of the invention, the invention includes anisolated protein fragment comprising a binding pocket or active sitedefined by structure coordinates of HDM2 amino acid residues Ser¹⁷,Ile¹⁹, Leu⁸² and Arg⁹⁷. In one embodiment, the isolated fragment islinked to a solid support.

[0092] In another aspect of the invention, the invention includes anisolated nucleic acid molecule encoding the fragment which comprises abinding pocket or active site defined by structure coordinates of HDM2amino acid residues Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷. In one embodiment, avector comprises the nucleic acid molecule. In another embodiment, ahost cell comprises the vector. In yet another aspect of the invention,the invention includes a method of producing a protein fragment,comprising culturing the host cell under conditions in which thefragment is expressed. In another aspect of the invention, the inventionincludes a method of screening for an agent that associates with HDM2,comprising: (a) exposing a protein molecule fragment to the agent; and(b) detecting the level of association of the agent to the fragment. Inanother aspect of the invention, the invention includes a kit comprisinga protein molecule fragment.

[0093] In another aspect of the invention, the invention includes amethod for the production of a crystal complex comprising an HDM2polypeptide-ligand comprising: (a) contacting the HDM2 polypeptide withsaid ligand in a suitable solution comprising PEG and NaSCN; and, b)crystallizing said resulting complex of HDM2 polypeptide-ligand fromsaid solution. In one embodiment, the HDM2 polypeptide is a polypeptidehaving SEQ ID NO: 2. In another embodiment, PEG has an average molecularweight range from 100 to 1000, wherein said PEG is present in solutionat a range from about 0.5% w/v to about 10% w/v and said NaSCN ispresent in solution at a range of from about 50 mM to about 150 mM. In apreferred embodiment, PEG has an average molecular weight of about 400and is present in solution at about 2% w/v and said NaSCN is present insolution at about 100 mM. In a highly preferred embodiment, the solutionfurther comprises about 1.8-2.4 M (NH₄)₂SO₄ and about 100 mM buffer.

[0094] The invention further includes a method for the production of acrystal comprising HDM2 and a ligand wherein the ligand is a smallmolecule inhibitor comprising crystallizing a peptide comprising asequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO: 3 and SEQ ID NO:4 with a potential inhibitor.

[0095] The invention includes a method for identifying a potentialinhibitor of HDM2 comprising: a) using a three dimensional structure ofHDM2 as defined by atomic coordinates according to table 1; b) replacingone or more HDM2 amino acids selected from Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷in said three-dimensional structure with a different amino acid toproduce a modified HDM2; c) using said three-dimensional structure todesign or select said potential inhibitor; d) synthesizing saidpotential inhibitor; and, e) contacting said potential inhibitor withsaid modified HDM2 in the presence of a substrate to test the ability ofsaid potential inhibitor to inhibit HDM2 or said modified HDM2. In oneembodiment, replacing one or more amino acid residues further comprisesreplacing SEQ ID NO: 2 amino acids selected from the group consisting ofVal⁵³, Leu⁵⁴, Phe⁵⁵, Leu⁵⁷, Gly⁵⁸, Gln⁵⁹, Ile⁶¹, Met⁶², Tyr⁶⁷, Gln⁷²,His⁷³, Ile⁷⁴, Val⁷⁵, Phe⁸⁶, Phe⁹¹, Val⁹³, Lys⁹⁴, Glu⁹⁵, His⁹⁶, Ile⁹⁹,Tyr¹⁰⁰, and Ile¹⁰³. In another embodiment, the potential inhibitor isselected from a database. In a preferred embodiment, the potentialinhibitor is designed de novo. In another preferred embodiment, thepotential inhibitor is designed from a known inhibitor. In a highlypreferred embodiment, the step of employing said three-dimensionalstructure to design or select said potential inhibitor comprises thesteps of: a) identifying chemical entities or fragments capable ofassociating with modified HDM2; and b) assembling the identifiedchemical entities or fragments into a single molecule to provide thestructure of said potential inhibitor. In one embodiment, the potentialinhibitor is a competitive inhibitor of SEQ ID NO:4 (Gly¹⁶-SEQ ID NO:2). In a different embodiment, the potential inhibitor is anon-competitive or uncompetitive inhibitor of SEQ ID NO:4 (Gly¹⁶-SEQ IDNO: 2). In yet another embodiment, an inhibitor is identified by themethod.

[0096] A. Modeling the Three-Dimensional Structure of HDM2

[0097] The atomic coordinate data provided in Table 1, Table 2 or thecoordinate data derived from homologous proteins may be used to build athree-dimensional model of HDM2. Any available computational methods maybe used to build the three dimensional model. As a starting point, theX-ray diffraction pattern obtained from the assemblage of the moleculesor atoms in a crystalline version of HDM2 or an HDM2 homolog can be usedto build an electron density map using tools well known to those skilledin the art of crystallography and X-ray diffraction techniques.Additional phase information extracted either from the diffraction dataand available in the published literature and/or from supplementingexperiments may then used to complete the reconstruction.

[0098] For basic concepts and procedures of collecting, analyzing, andutilizing X-ray diffraction data for the construction of electrondensities see, for example, Campbell et al., 1984, BiologicalSpectroscopy, The Benjamin/Cummings Publishing Co., Inc., Menlo Park,Calif.; Cantor et al., 1980, Biophysical Chemistry, Part II: Techniquesfor the study of biological structure and function, W. H. Freeman andCo., San Francisco, Calif.; A. T. Brunger, 1993, X-Flor Version 3.1: Asystem for X-ray crystallography and NMR, Yale Univ. Pr., New Haven,Conn.; M. M. Woolfson, 1997, An Introduction to X-ray Crystallography,Cambridge Univ. Pr., Cambridge, UK; J. Drenth, 1999, Principles ofProtein X-ray Crystallography (Springer Advanced Texts in Chemistry),Springer Verlag; Berlin; Tsirelson et al., 1996, Electron Density andBonding in Crystals: Principles, Theory and X-ray DiffractionExperiments in Solid State Physics and Chemistry, Inst. of Physics Pub.;U.S. Pat. No. 5,942,428; U.S. Pat. No. 6,037,117; U.S. Pat. No.5,200,910 and U.S. Pat. No. 5,365,456 (“Method for Modeling the ElectronDensity of a Crystal”), each of which is herein specificallyincorporated by reference in their entirety.

[0099] For basic information on molecular modeling, see, for example, M.Schlecht, Molecular Modeling on the PC, 1998, John Wiley & Sons; Gans etal., Fundamental Principals of Molecular Modeling, 1996, Plenum Pub.Corp.; N. C. Cohen (editor), Guidebook on Molecular Modeling in DrugDesign, 1996, Academic Press; and W. B. Smith, Introduction toTheoretical Organic Chemistry and Molecular Modeling, 1996. U.S. Patentswhich provide detailed information on molecular modeling include U.S.Pat. Nos. 6,093,573; 6,080,576; 6,075,014; 6,075,123; 6,071,700;5,994,503; 5,612,894; 5,583,973; 5,030,103; 4,906,122; and 4,812,12,each of which are incorporated by reference herein in their entirety.

[0100] B. Methods of Using the Atomic Coordinates to Identify and DesignLigands of Interest

[0101] The atomic coordinates of the invention, such as those describedin Table 1, Table 2, Table 3 or coordinates substantially identical toor homologous to those of Table 1, Table 2, or Table 3 may be used withany available methods to prepare three dimensional models of HDM2 aswell as to identify and design HDM2 ligands, inhibitors or antagonistsor agonist molecules.

[0102] For instance, three-dimensional modeling may be performed usingthe experimentally determined coordinates derived from X-ray diffractionpatterns, such as those in Table 1 or Table 2, for example, wherein suchmodeling includes, but is not limited to, drawing pictures of the actualstructures, building physical models of the actual structures, anddetermining the structures of related subunits and HDM2/ligand and HDM2subunit/ligand complexes using the coordinates. Such molecular modelingcan utilize known X-ray diffraction molecular modeling algorithms ormolecular modeling software to generate atomic coordinates correspondingto the three-dimensional structure of HDM2.

[0103] As described above, molecular modeling involves the use ofcomputational methods, preferably computer assisted methods, to buildrealistic models of molecules that are identifiably related in sequenceto the known crystal structure. It also involves modeling new smallmolecule inhibitors bound to HDM2 starting with the structures of HDM2and or HDM2 complexed with known ligands or inhibitors. The methodsutilized in ligand modeling range from molecular graphics (i.e., 3Drepresentations) to computational chemistry (i.e., calculations of thephysical and chemical properties) to make predictions about the bindingof ligands or activities of ligands; to design new ligands; and topredict novel molecules, including ligands such as drugs, for chemicalsynthesis, collectively referred to as rational drug design.

[0104] One approach to rational drug design is to search for knownmolecular structures that might bind to an active site. Using molecularmodeling, rational drug design programs can look at a range of differentmolecular structures of drugs that may fit into the active site of anenzyme, and by moving them in a three-dimensional environment it can bedecided which structures actually fit the site well. See, for example,U.S. Appl. Nos. 60/275,629; 60/331,235; 60/379,617; and, 10/097,249.See, also, for example, data in Tables 1, 2 and 3.

[0105] An alternative but related rational drug design approach startswith the known structure of a complex with a small molecule ligand andmodels modifications of that small molecule in an effort to makeadditional favorable interactions with HDM2.

[0106] The present invention include the use of molecular and computermodeling techniques to design and select and design ligands, such assmall molecule agonists or antagonists or other therapeutic agents thatinteract with HDM2. Such agents include, but are not limited to 1,4benzodiazepines and derivatives thereof. For example, the invention asherein described includes the design of ligands that act as competitiveinhibitors of at least one HDM2 function by binding to all, or a portionof, the active sites or other regions of HDM2.

[0107] This invention also includes the design of compounds that act asuncompetitive inhibitors of at least one function of HDM2. Theseinhibitors may bind to all, or a portion of, the active sites or otherregions of HDM2 already bound to its substrate and may be more potentand less non-specific than competitive inhibitors that compete for HDM2active sites. Similarly, non-competitive inhibitors that bind to andinhibit at least one function of HDM2 whether or not it is bound toanother chemical entity may be designed using the atomic coordinates ofHDM2 or complexes comprising HDM2 of this invention.

[0108] The atomic coordinates of the present invention also provide theneeded information to probe a crystal of HDM2 with molecules composed ofa variety of different chemical features to determine optimal sites forinteraction between candidate inhibitors and/or activators and HDM2. Forexample, high resolution X-ray diffraction data collected from crystalssaturated with solvent allows the determination of where each type ofsolvent molecule sticks. Small molecules that bind to those sites canthen be designed and synthesized and tested for their inhibitoryactivity (Travis, J., Science 262:1374(1993)).

[0109] The present invention also includes methods for computationallyscreening small molecule databases and libraries for chemical entities,agents, ligands, or compounds that can bind in whole, or in part, toHDM2. In this screening, the quality of fit of such entities orcompounds to the binding site or sites may be judged either by shapecomplementarity or by estimated interaction energy (Meng, E. C. et al.,J. Coma. Chem. 13:505-524 (1992)).

[0110] The design of compounds that bind to promote or inhibit thefunctional activity of HDM2 according to this invention generallyinvolves consideration of two factors. First, the compound must becapable of physically and structurally associating with HDM2.Non-covalent molecular interactions important in the association of HDM2with the compound, include hydrogen bonding, van der Waals andhydrophobic interactions. Second, the compound must be able to assume aconformation that allows it to associate with HDM2. Although certainportions of the compound may not directly participate in the associationwith HDM2, those portions may still influence the overall conformationof the molecule. This, in turn, may have a significant impact on bindingaffinities, therapeutic efficacy, drug-like qualities and potency. Suchconformational requirements include the overall three-dimensionalstructure and orientation of the chemical entity or compound in relationto all or a portion of the active site or other region of HDM2, or thespacing between functional groups of a compound comprising severalchemical entities that directly interact with HDM2.

[0111] The potential, predicted, inhibitory agonist, antagonist orbinding effect of a ligand or other compound on HDM2 may be analyzedprior to its actual synthesis and testing by the use of computermodeling techniques. If the theoretical structure of the given compoundsuggests insufficient interaction and association between it and HDM2,synthesis and testing of the compound may be obviated. However, ifcomputer modeling indicates a strong interaction, the molecule may thenbe synthesized and tested for its ability to interact with HDM2. In thismanner, synthesis of inoperative compounds may be avoided. In somecases, inactive compounds are synthesized predicted on modeling and thentested to develop a SAR (structure-activity relationship) for compoundsinteracting with a specific region of HDM2.

[0112] One skilled in the art may use one of several methods to screenchemical entities fragments, compounds, or agents for their ability toassociate with HDM2 and more particularly with the individual bindingpockets or active sites of HDM2. This process may begin by visualinspection of, for example, the active site on the computer screen basedon the atomic coordinates of HDM2 or HDM2 complexed with a ligand.Selected chemical entities, compounds, or agents may then be positionedin a variety of orientations, or docked within an individual bindingpocket of HDM2. Docking may be accomplished using software such asQuanta and Sybyl, followed by energy minimization and molecular dynamicswith standard molecular mechanics forcefields, such as CHARMM and AMBER.

[0113] Specialized computer programs may also assist in the process ofselecting chemical entities. These include but are not limited to: GRID(Goodford, P. J., “A Computational Procedure for DeterminingEnergetically Favorable Binding Sites on Biologically ImportantMacromolecules,” J. Med. Chem. 28:849-857 (1985), available from OxfordUniversity, Oxford, UK); MCSS (Miranker, A. and M. Karplus,“Functionality Maps of Binding Sites: A Multiple Copy SimultaneousSearch Method.” Proteins: Structure, Function and Genetics 11: 29-34(1991), available from Molecular Simulations, Burlington, Mass.);AUTODOCK (Goodsell, D. S. and A. J. Olsen, “Automated Docking ofSubstrates to Proteins by Simulated Annealing” Proteins: Structure.Function, and Genetics 8:195-202 (1990), available from Scripps ResearchInstitute, La Jolla, Calif.); and DOCK (Kuntz, I. D. et al., “AGeometric Approach to Macromolecule-Ligand Interactions,” J.-Mol. Biol.161:269-288 (1982), available from University of California, SanFrancisco, Calif.).

[0114] The use of software such as GRID, a program that determinesprobable interaction sites between probes with various functional groupcharacteristics and the macromolecular surface, is used to analyze thesurface sites to determine structures of similar inhibiting proteins orcompounds. The GRID calculations, with suitable inhibiting groups onmolecules (e.g., protonated primary amines) as the probe, are used toidentify potential hotspots around accessible positions at suitableenergy contour levels. The program DOCK may be used to analyze an activesite or ligand binding site and suggest ligands with complementarysteric properties.

[0115] Once suitable chemical entities, compounds, or agents have beenselected, they can be assembled into a single ligand or compound orinhibitor or activator. Assembly may proceed by visual inspection of therelationship of the fragments to each other on the three-dimensionalimage. This may be followed by manual model building using software suchas Quanta or Sybyl.

[0116] Useful programs to aid in connecting the individual chemicalentities, compounds, or agents include but are not limited to: CAVEAT(Bartlett, P. A. et al., “CAVEAT: A Program to Facilitate theStructure-Derived Design of Biologically Active Molecules.” In MolecularRecognition in Chemical and Biological Problems, Special Pub., RoyalChem. Soc., 78, pp. 82-196 (1989)); 3D Database systems such as MACCS-3D(MDL Information Systems, San Leandro, Calif. and Martin, Y.C., “3DDatabase Searching in Drug Design”, J. Med. Chem. 35: 2145-2154 (1992);and HOOK (available from Molecular Simulations, Burlington, Mass.).

[0117] Several methodologies for searching three-dimensional databasesto test pharmacophore hypotheses and select compounds for screening areavailable. These include the program CAVEAT (Bacon et al., J. Mol. Biol.225:849-858 (1992)). For instance, CAVEAT uses databases of cycliccompounds which can act as “spacers” to connect any number of chemicalfragments already positioned in the active site. This allows one skilledin the art to quickly generate hundreds of possible ways to connect thefragments already known or suspected to be necessary for tight binding.

[0118] Instead of proceeding to build an inhibitor activator, agonist orantagonist of HDM2 in a step-wise fashion one chemical entity at a timeas described above, such compounds may be designed as a whole or “denovo” using either an empty active site or optionally including someportion(s) of a known molecules. These methods include: LUDI (Bohm,H.-J., “The Computer Program LUDI: A New Method for the De Novo Designof Enzyme Inhibitors”, J. ComR. Aid. Molec. Design, 6, pp. 61-78 (1992),available from Biosym Technologies, San Diego, Calif.); LEGEND(Nishibata, Y. and A. Itai, Tetrahedron 47:8985 (1991), available fromMolecular Simulations, Burlington, Mass.); and LeapFrog (available fromTripos Associates, St. Louis, Mo.).

[0119] For instance, the program LUDI can determine a list ofinteraction sites into which to place both hydrogen bonding andhydrophobic fragments. LUDI then uses a library of linkers to connect upto four different interaction sites into fragments. Then smaller“bridging” groups such as —CH2- and —COO— are used to connect thesefragments. For example, for the enzyme DHFR, the placements of keyfunctional groups in the well-known inhibitor methotrexate werereproduced by LUDI. See also, Rotstein and Murcko, J. Med. Chem. 36:1700-1710 (1992).

[0120] Other molecular modeling techniques may also be employed inaccordance with this invention. See, e.g., Cohen, N. C. et al.,“Molecular Modeling Software and Methods for Medicinal Chemistry, J.Med. Chem. 33:883-894 (1990). See also, Navia, M. A. and M. A. Murcko,“The Use of Structural Information in Drug Design,” Current Opinions inStructural Biology, 2, pp. 202-210 (1992).

[0121] Once a compound has been designed or selected by the abovemethods, the affinity with which that compound may bind or associatewith HDM2 may be tested and optimized by computational evaluation and/orby testing biological activity after synthesizing the compound.Inhibitors or compounds may interact with the HDM2 in more than oneconformation that is similar in overall binding energy. In those cases,the deformation energy of binding is taken to be the difference betweenthe energy of the free compound and the average energy of theconformations observed when the compound binds to HDM2.

[0122] A compound designed or selected as binding or associating withHDM2 may be further computationally optimized so that in its bound stateit would preferably lack repulsive electrostatic interaction with HDM2.Such non-complementary (e.g., electrostatic) interactions includerepulsive charge-charge, dipole-dipole and charge-dipole interactions.Specifically, the sum of all electrostatic interactions between theinhibitor and HDM2 when the inhibitor is bound, preferably make aneutral or favorable contribution to the enthalpy of binding. Weakbinding compounds will also be designed by these methods so as todetermine SAR. See, for example, U.S. Appl. Nos. 60/275,629; 60/331,235;60/379,617; and, 10/097,249.

[0123] Specific computer software is available in the art to evaluatecompound deformation energy and electrostatic interaction. Examples ofprograms designed for such uses include: Gaussian 92, revision C (M. J.Frisch, Gaussian, Inc., Pittsburgh, Pa., COPYRGT 1992); AMBER, version4.0 (P. A. Kollman, University of California at San Francisco, COPYRGT1994); QUANTA/CHARMM (Molecular Simulations, Inc., Burlington, Mass.COPYRGT 1994); and Insight II/Discover (Biosysm Technologies Inc., SanDiego, Calif. COPYRGT 1994). Other hardware systems and softwarepackages will be known to those skilled in the art.

[0124] Once a compound that associates with HDM2 has been optimallyselected or designed, as described above, substitutions may then be madein some of its atoms or side groups in order to improve or modify itsbinding properties. Generally, initial substitutions are conservative,i.e., the replacement group will have approximately the same size,shape, hydrophobicity and charge as the original group. It should, ofcourse, be understood that components known in the art to alterconformation may be avoided. Such substituted chemical compounds maythen be analyzed for efficiency of fit to HDM2 by the same computermethods described in detail, above.

[0125] C. Use of Homology Structure Modeling to Design Ligands withModulated Binding or Activity to HDM2

[0126] The present invention includes the use of the atomic coordinatesand structures of HDM2 and/or HDM2 complexed with an inhibitor to designmodifications to starting compounds, such as(4-Choloro-phenyl)-[3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-aceticacid;[8-Chloro-3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-(4-chloro-phenyl)-aceticacid; and derivatives thereof that will bind more tightly or interactmore specifically to the target enzyme. See, U.S. Appl. Nos. 60/275,629;60/331,235; 60/379,617; and, 10/097,249, disclosing compounds 1 and 2and derivatives thereof, all of which are incorporated herein in theirentirety.

[0127] Compound 1 (338437):(4-Chloro-phenyl)-[3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-aceticacid

[0128] Compound 2 (876273):[8-Chloro-3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-(4-chloro-phenyl)-aceticacid

[0129] The structure of a complex between the HDM2 and the startingcompound can be used to guide the modification of that compound toproduce new compounds that have other desirable properties forapplicable industrial and other uses (e.g., as pharmaceuticals), such aschemical stability, solubility or membrane permeability. (Lipinski etal., Adv. Drug Deliv. Rev. 23:3 (1997)).

[0130] Binding compounds, agonists, antagonists and such that are knownin the art include but are not limited to p53 peptides and smallmolecule antagonists. See, for example, U.S. Appl. Nos. 60/275,629;60/331,235; 60/379,617; and, 10/097,249 incorporated by reference hereinin their entirety. Such compounds can be diffused into or soaked withthe stabilized crystals of HDM2 to form a complex for collecting X-raydiffraction data. Alternatively, the compounds, known and unknown in theart, can be cocrystallized with HDM2 by mixing the compound with HDM2before precipitation.

[0131] To produce custom high affinity and very specific compounds, thestructure of HDM2 can be compared to the structure of a selectednon-targeted molecule and a hybrid constructed by changing the structureof residues at the binding site for a ligand for the residues at thesame positions of the non-target molecule. The process whereby thismodeling is achieved is referred to as homology structure modeling. Thisis done computationally by removing the side chains from the molecule ortarget of known structure and replacing them with the side chains of theunknown structure put in sterically plausible positions. In this way itcan be understood how the shapes of the active site cavities of thetargeted and non-targeted molecules differ. This process, therefore,provides information concerning how a bound ligand can be chemicallyaltered in order to produce compounds that will bind tightly andspecifically to the desired target but will simultaneously be stericallyprevented from binding to the non-targeted molecule. Likewise, knowledgeof portions of the bound ligands that are facing to the solvent wouldallow introduction of other functional groups for additionalpharmaceutical purposes. The use of homology structure modeling todesign molecules (ligands) that bind more tightly to the target enzymethan to the non-target enzyme has wide spread applicability.

[0132] D. High Throughput Assays

[0133] Any high throughput screening may be utilized to test newcompounds which are identified or designed for their ability to interactwith HDM2. For general information on high-throughput screening see, forexample, Devlin, 1998, High Throughput Screening, Marcel Dekker; andU.S. Pat. No. 5,763,263. High throughput assays utilize one or moredifferent assay techniques including, but not limited to, thosedescribed below.

[0134] Immunodiagnostics and Immunoassays. These are a group oftechniques used for the measurement of specific biochemical substances,commonly at low concentrations in complex mixtures such as biologicalfluids, that depend upon the specificity and high affinity shown bysuitably prepared and selected antibodies for their complementaryantigens. A substance to be measured must, of necessity, beantigenic—either an immunogenic macromolecule or a haptenic smallmolecule. To each sample a known, limited amount of specific antibody isadded and the fraction of the antigen combining with it, often expressedas the bound:free ratio, is estimated, using as indicator a form of theantigen labeled with radioisotope (radioimmunoassay), fluorescentmolecule (fluoroimmunoassay), stable free radical (spin immunoassay),enzyme (enzyme immunoassay), or other readily distinguishable label.

[0135] Antibodies can be labeled in various ways, including:enzyme-linked immunosorbent assay (ELISA); radioimmuno assay (RIA);fluorescent immunoassay (FIA); chemiluminescent immunoassay (CLIA); andlabeling the antibody with colloidal gold particles (immunogold).

[0136] Common assay formats include the sandwich assay, competitive orcompetition assay, latex agglutination assay, homogeneous assay,microtitre plate format and the microparticle-based assay.

[0137] Enzyme-linked immunosorbent assay (ELISA). ELISA is animmunochemical technique that avoids the hazards of radiochemicals andthe expense of fluorescence detection systems. Instead, the assay usesenzymes as indicators. ELISA is a form of quantitative immunoassay basedon the use of antibodies (or antigens) that are linked to an insolublecarrier surface, which is then used to “capture” the relevant antigen(or antibody) in the test solution. The antigen-antibody complex is thendetected by measuring the activity of an appropriate enzyme that hadpreviously been covalently attached to the antigen (or antibody).

[0138] For information on ELISA techniques, see, for example, Crowther,(1995) ELISA—Theory and Practice (Methods in Molecular Biology), HumanaPress; Challacombe & Kemeny, (1998) ELISA and Other Solid PhaseImmunoassays—Theoretical and Practical Aspects, John Wiley; Kemeny,(1991) A Practical Guide to ELISA, Pergamon Press; Ishikawa, (1991)Ultrasensitive and Rapid Enzyme Immunoassay (Laboratory Techniques inBiochemistry and Molecular Biology) Elsevier.

[0139] Colorimetric Assays for Enzymes. Colorimetry is any method ofquantitative chemical analysis in which the concentration or amount of acompound is determined by comparing the color produced by the reactionof a reagent with both standard and test amounts of the compound, oftenusing a colorimeter. A colorimeter is a device for measuring colorintensity or differences in color intensity, either visually orphotoelectrically.

[0140] Standard colorimetric assays of beta-galactosidase enzymaticactivity are well known to those skilled in the art (see, for example,Norton et al., Mol. Cell. Biol. 5:281-290 (1985). A colorimetric assaycan be performed on whole cell lysates usingO-nitrophenyl-beta-D-galactopyranoside (ONPG, Sigma) as the substrate ina standard colorimetric beta-galactosidase assay (Sambrook et al.,(1989) Molecular Cloning—A Laboratory Manual, Cold Spring HarborLaboratory Press). Automated colorimetric assays are also available forthe detection of beta-galactosidase activity, as described in U.S. Pat.No. 5,733,720.

[0141] Immunofluorescence Assays. Immunofluorescence orimmunofluorescence microscopy is a technique in which an antigen orantibody is made fluorescent by conjugation to a fluorescent dye andthen allowed to react with the complementary antibody or antigen in atissue section or smear. The location of the antigen or antibody canthen be determined by observing the fluorescence by microscopy underultraviolet light.

[0142] For general information on immunofluorescent techniques, see, forexample, Knapp et al., (1978) Immunofluorescence and Related StainingTechniques, Elsevier; Allan, (1999) Protein Localization by FluorescentMicroscopy—A Practical Approach (The Practical Approach Series) OxfordUniversity Press; Caul, (1993) Immunofluorescence Antigen DetectionTechniques in Diagnostic Microbiology, Cambridge University Press. Fordetailed explanations of immunofluorescent techniques applicable to thepresent invention, see U.S. Pat. No. 5,912,176; U.S. Pat. No. 5,869,264;U.S. Pat. No. 5,866,319; and U.S. Pat. No. 5,861,259.

[0143] E. Databases and Computer Systems

[0144] An amino acid sequence or nucleotide sequence of HDM2 and/orX-ray diffraction data, useful for computer molecular modeling of HDM2or a portion thereof, can be “provided” in a variety of mediums tofacilitate use thereof. As used herein, “provided” refers to amanufacture, which contains, for example, an amino acid sequence ornucleotide sequence and/or atomic coordinates derived from X-raydiffraction data of the present invention, e.g., an amino acid ornucleotide sequence of HDM2, a representative fragment thereof, or ahomologue thereof. Such a method provides the amino acid sequence and/orX-ray diffraction data in a form which allows a skilled artisan toanalyze and molecular model the three-dimensional structure of HDM2 orrelated molecules, including a subdomain thereof.

[0145] In one application of this embodiment, databases comprising datapertaining to HDM2, or at least one subdomain thereof, amino acid andnucleic acid sequence and/or X-ray diffraction data of the presentinvention is recorded on computer readable medium. As used herein,“computer readable medium” refers to any medium which can be read andaccessed directly by a computer. Such media include, but are not limitedto: magnetic storage media, such as floppy discs, hard disc storagemedia, and magnetic tape; optical storage media such as optical discs orCD-ROM; electrical storage media such as RAM and ROM; and hybrids ofthese categories such as magnetic/optical storage media. A skilledartisan can readily appreciate how any of the presently known computerreadable media can be used to create a manufacture comprising computerreadable medium having recorded thereon an amino acid sequence and/orX-ray diffraction data of the present invention.

[0146] As used herein, “recorded” refers to a process for storinginformation on computer readable media. A skilled artisan can readilyadopt any of the presently known methods for recording information oncomputer readable media to generate manufactures comprising an aminoacid sequence and/or atomic coordinate/X-ray diffraction datainformation of the present invention.

[0147] A variety of data storage structures are available to a skilledartisan for creating a computer readable medium having recorded thereonan amino acid sequence and/or atomic coordinate/X-ray diffraction dataof the present invention. The choice of the data storage structure willgenerally be based on the means chosen to access the stored information.In addition, a variety of data processor programs and formats can beused to store the sequence and X-ray data information of the presentinvention on computer readable media. The sequence information can berepresented in a word processing text file, formatted incommercially-available software such as WordPerfect and MICROSOFT Word,or represented in the form of an ASCII file, stored in a databaseapplication, such as DB2, Sybase, Oracle, or the like. A skilled artisancan readily adapt any number of dataprocessor structuring formats (e.g.text file or database) in order to obtain computer readable media havingrecorded thereon the information of the present invention.

[0148] By providing computer readable media having sequence and/oratomic coordinates based on X-ray diffraction data, a skilled artisancan routinely access the sequence and atomic coordinate or X-raydiffraction data to model a related molecule, a subdomain, mimetic, or aligand thereof. Computer algorithms are publicly and commerciallyavailable which allow a skilled artisan to access this data provided ina computer readable medium and analyze it for molecular modeling and/orRDD (rational drug design). See, e.g., Biotechnology Software Directory,MaryAnn Liebert Publ., New York (1995).

[0149] The present invention further provides systems, particularlycomputer-based systems, which contain the sequence and/or diffractiondata described herein. Such systems are designed to do structuredetermination and RDD for HDM2 or at least one subdomain thereof.Non-limiting examples are microcomputer workstations available fromSilicon Graphics Incorporated and Sun Microsystems running UNIX based,Windows NT or IBM OS/2 operating systems.

[0150] As used herein, “a computer-based system” refers to the hardwaremeans, software means, and data storage means used to analyze thesequence and/or X-ray diffraction data of the present invention. Theminimum hardware means of the computer-based systems of the presentinvention comprises a central processing unit (CPU), input means, outputmeans, and data storage means. A skilled artisan can readily appreciatewhich of the currently available computer-based systems are suitable foruse in the present invention. A visualization device, such as a monitor,is optionally provided to visualize structure data.

[0151] As stated above, the computer-based systems of the presentinvention comprise a data storage means having stored therein sequenceand/or atomic coordinate/X-ray diffraction data of the present inventionand the necessary hardware means and software means for supporting andimplementing an analysis means. As used herein, “data storage means”refers to memory which can store sequence or atomic coordinate/X-raydiffraction data of the present invention, or a memory access meanswhich can access manufactures having recorded thereon the sequence orX-ray data of the present invention.

[0152] As used herein, “search means” or “analysis means” refers to oneor more programs which are implemented on the computer-based system tocompare a target sequence or target structural motif with the sequenceor X-ray data stored within the data storage means. Search means areused to identify fragments or regions of a protein which match aparticular target sequence or target motif. A variety of knownalgorithms are disclosed publicly and a variety of commerciallyavailable software for conducting search means are and can be used inthe computer-based systems of the present invention. A skilled artisancan readily recognize that any one of the available algorithms orimplementing software packages for conducting computer analyses can beadapted for use in the present computer-based systems.

[0153] As used herein, “a target structural motif,” or “target motif,”refers to any rationally selected sequence or combination of sequencesin which the sequence(s) are chosen based on a three-dimensionalconfiguration or electron density map which is formed upon the foldingof the target motif. There are a variety of target motifs known in theart. Protein target motifs include, but are not limited to, enzymaticactive sites, inhibitor binding sites, structural subdomains, epitopes,functional domains and signal sequences. Similar motifs are known forRNA. A variety of structural formats for the input and output means canbe used to input and output the information in the computer-basedsystems of the present invention.

[0154] A variety of comparing means can be used to compare a targetsequence or target motif with the data storage means to identifystructural motifs or electron density maps derived in part from theatomic coordinate/X-ray diffraction data. A skilled artisan can readilyrecognize that any one of the publicly available computer modelingprograms can be used as the search means for the computer-based systemsof the present invention.

[0155] F. Target Molecule Fragments and Portions

[0156] Fragments of HDM2, for instance fragments comprising active sitesdefined by two or more amino acids selected from the group consistingof: Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷, may be prepared by any availablemeans including synthetic or recombinant means. Such fragments may thenbe used in the assays as described above, for instance, high through-putassays to detect interactions between prospective agents and the activesite within the fragment.

[0157] For recombinant expression or production of the fragments of theinvention, nucleic acid molecules encoding the fragment may be prepared.As used herein, “nucleic acid” is defined as RNA or DNA that encodes aprotein or peptide as defined above, or is complementary to nucleic acidsequence encoding such peptides, or hybridizes to such nucleic acid andremains stably bound to it under appropriate stringency conditions.

[0158] Nucleic acid molecules encoding fragments of the invention maydiffer in sequence because of the degeneracy in the genetic code or maydiffer in sequence as they encode proteins or protein fragments thatdiffer in amino acid sequence. Homology or sequence identity between twoor more such nucleic acid molecules is determined by BLAST (Basic LocalAlignment Search Tool) analysis using the algorithm employed by theprograms blastp, blastn, blastx, tblastn and tblastx (Karlin et al.,Proc. Natl. Acad. Sci. USA 87:2264-2268 (1990) and Altschul, et al., J.Mol. Evol. 36:290-300 (1993), fully incorporated by reference) which aretailored for sequence similarity searching.

[0159] The approach used by the BLAST program is to first considersimilar segments between a query sequence and a database sequence, thento evaluate the statistical significance of all matches that areidentified and finally to summarize only those matches which satisfy apreselected threshold of significance. For a discussion of basic issuesin similarity searching of sequence databases, see Altschul et al. (Nat.Genet. 6, 119-129 (1994)) which is fully incorporated by reference. Thesearch parameters for histogram, descriptions, alignments, expect (i.e.,the statistical significance threshold for reporting matches againstdatabase sequences), cutoff, matrix and filter are at the defaultsettings. The default scoring matrix used by blastp, blastx, tblastn,and tblastx is the BLOSUM62 matrix (Henikoff et al., Proc. Natl. Acad.Sci. USA 89:10915-10919 (1992), fully incorporated by reference). Fourblastn parameters were adjusted as follows: Q=10 (gap creation penalty);R=10 (gap extension penalty); wink=1 (generates word hits at everywink^(th) position along the query); and gapw=16 (sets the window widthwithin which gapped alignments are generated). The equivalent Blastpparameter settings were Q=9; R=2; wink=1; and gapw=32. A Bestfitcomparison between sequences, available in the GCG package version 10.0,uses DNA parameters GAP=50 (gap creation penalty) and LEN=3 (gapextension penalty) and the equivalent settings in protein comparisonsare GAP=8 and LEN=2.

[0160] “Stringent conditions” are those that (1) employ low ionicstrength and high temperature for washing, for example, 0.015 MNaCl/0.0015 M sodium citrate/0.1% SDS at 50° C. or (2) employ duringhybridization a denaturing agent such as formamide, for example, 50%formamide with 0.1% bovine serum albumin/0.1% Ficoll/0.1%polyvinylpyrrolidone/50 mM sodium phosphate buffer at pH 6.5 with 750 mMNaCl, 75 mM sodium citrate at 42° C. Another example is use of 50%formamide, 5×SSC, 50 mM sodium phosphate (pH 6.8), 0.1% sodiumpyrophosphate, 5× Denhardt's solution, sonicated salmon sperm DNA (50mg/ml), 0.1% SDS and 10% dextran sulfate at 42° C., with washes at 42°C. in 0.2×SSC and 0.1% SDS. A skilled artisan can readily determine andvary the stringency conditions appropriately to obtain a clear anddetectable hybridization signal.

[0161] As used herein, a nucleic acid molecule is said to be “isolated”when the nucleic acid molecule is substantially separated fromcontaminant nucleic acid encoding other polypeptides from the source ofnucleic acid.

[0162] The encoding nucleic acid molecules of the present invention(i.e., synthetic oligonucleotides) and those that are used as probes orspecific primers for polymerase chain reaction (PCR) or to synthesizegene sequences encoding proteins of the invention can easily besynthesized by chemical techniques, for example, the phosphotriestermethod of Matteucci et al. (J. Am. Chem. Soc. 103: 185-3191 (1981)) orusing automated synthesis methods. In addition, larger DNA segments canreadily be prepared by well known methods, such as synthesis of a groupof oligonucleotides that define various modular segments of the gene,followed by ligation of oligonucleotides to build the complete modifiedgene.

[0163] The encoding nucleic acid molecules of the present invention mayfurther be modified so as to contain a detectable label for diagnosticand probe purposes. A variety of such labels are known in the art andcan readily be employed with the encoding molecules herein described.Suitable labels include, but are not limited to, biotin, radiolabelednucleotides and the like. A skilled artisan can employ any of theart-known labels to obtain a labeled encoding nucleic acid molecule.

[0164] The present invention further provides recombinant DNA molecules(rDNA) that contain a coding sequence for a protein fragment asdescribed above. As used herein, a rDNA molecule is a DNA molecule thathas been subjected to molecular manipulation. Methods for generatingrDNA molecules are well known in the art, for example, see Sambrook etal. Molecular Cloning—A Laboratory Manual, Cold Spring Harbor LaboratoryPress (1989). In the preferred rDNA molecules, a coding DNA sequence isoperably linked to expression control sequences and/or vector sequences.

[0165] The choice of vector and expression control sequences to whichone of the protein encoding sequences of the present invention isoperably linked depends directly, as is well known in the art, on thefunctional properties desired (e.g., protein expression, and the hostcell to be transformed). A vector of the present invention may becapable of directing the replication or insertion into the hostchromosome, and preferably also expression, of the structural geneincluded in the rDNA molecule.

[0166] Expression control elements that are used for regulating theexpression of an operably linked protein encoding sequence are known inthe art and include, but are not limited to, inducible promoters,constitutive promoters, secretion signals, and other regulatoryelements. Preferably, the inducible promoter is readily controlled, suchas being responsive to a nutrient in the host cell's medium.

[0167] The present invention further provides host cells transformedwith a nucleic acid molecule that encodes a protein fragment of thepresent invention. The host cell can be either prokaryotic oreukaryotic. Eukaryotic cells useful for expression of a protein of theinvention are not limited, so long as the cell line is compatible withcell culture methods and compatible with the propagation of theexpression vector and expression of the gene product. Preferredeukaryotic host cells include, but are not limited to, yeast, insect andmammalian cells, preferably vertebrate cells such as those from a mouse,rat, monkey or human cell line. Preferred eukaryotic host cells includeChinese hamster ovary (CHO) cells available from the ATCC as CCL61, NIHSwiss mouse embryo cells NIH-3T3 available from the ATCC as CRL1658,baby hamster kidney cells (BHK), and the like eukaryotic tissue culturecell lines.

[0168] Transformed host cells of the invention may be cultured underconditions that allow the production of the recombinant protein.Optionally the recombinant protein is isolated from the medium or fromthe cells; recovery and purification of the protein may not be necessaryin some instances where some impurities may be tolerated.

[0169] Kits may also be prepared with any of the above described nucleicacid molecules, protein fragments, vector and/or host cells optionallypackaged with the reagents needed for a specific assay, such as thosedescribed above. In such kits, the protein fragments or other reagentsmay be attached to a solid support, such as glass or plastic beads.

[0170] G. Integrated Procedures Which Utilize the Present Invention

[0171] Molecular modeling is provided by the present invention forrational drug design (RDD) of mimetics and ligands of HDM2. As describedabove, the drug design paradigm uses computer modeling programs todetermine potential mimetics and ligands which are expected to interactwith sites on the protein. The potential mimetics or ligands are thenscreened for activity and/or binding and/or interaction. ForHDM2-related mimetics or ligands, screening methods can be selected fromassays for at least one biological activity of HDM2, e.g., such asblocking p53 binding, according to known method steps. See, for example,Kussie et al., Science 274:948-953 (1996); Bottger et al., J. Mol. Biol.269:744-756 (1997).

[0172] Thus, the tools and methodologies provided by the presentinvention may be used in procedures for identifying and designingligands which bind in desirable ways with the target. Such proceduresutilize an iterative process whereby ligands are synthesized, tested andcharacterized. New ligands can be designed based on the informationgained in the testing and characterization of the initial ligands andthen such newly identified ligands can themselves be tested andcharacterized. This series of processes may be repeated as many times asnecessary to obtain ligands with the desirable binding properties.

[0173] The following steps serve as an example of the overall procedure:

[0174] 1. A biological activity of a target is selected (e.g., bindingto p53).

[0175] 2. A ligand is identified that appears to be in some wayassociated with the chosen biological activity (e.g., the ligand may bean inhibitor of a known activity). The activity of the ligand may betested by in vivo and/or in vitro methods.

[0176] A ligand of the present invention can be, but is not limited to,at least one selected from a lipid, a nucleic acid, a compound, aprotein, an element, an antibody, a saccharide, an isotope, acarbohydrate, an imaging agent, a lipoprotein, a glycoprotein, anenzyme, a detectable probe, and antibody or fragment thereof, or anycombination thereof, which can be detectably labeled as for labelingantibodies. Such labels include, but are not limited to, enzymaticlabels, radioisotope or radioactive compounds or elements, fluorescentcompounds or metals, chemiluminescent compounds and bioluminescentcompounds. Alternatively, any other known diagnostic or therapeuticagent can be used in a method of the invention. Suitable compounds arethen tested for activities in relationship to the target.

[0177] Complexes between HDM2 and ligands are made either byco-crystallization or more commonly by diffusing the small moleculeligand into the crystal. X-ray diffraction data from the complex crystalare measured and a difference electron density map is calculated. Thisprocess provides the precise location of the bound ligand on the targetmolecule. The difference Fourier is calculated using measure diffractionamplitudes and the phases of these reflections calculated from thecoordinates.

[0178] 3. Using the methods of the present invention, X-raycrystallography is utilized to create electron density maps and/ormolecular models of the interaction of the ligand with the targetmolecule.

[0179] The entry of the coordinates of the target into the computerprograms discussed above results in the calculation of most probablestructure of the macromolecule. These structures are combined andrefined by additional calculations using such programs to determine theprobable or actual three-dimensional structure of the target includingpotential or actual active or binding sites of ligands. Such molecularmodeling (and related) programs useful for rational drug design ofligands or mimetics, are also provided by the present invention.

[0180] 4. The electron density maps and/or molecular models obtained inStep 3 are compared to the electron density maps and/or molecular modelsof a non-ligand containing target and the observed/calculateddifferences are used to specifically locate the binding of the ligand onthe target or subunit.

[0181] 5. Modeling tools, such as computational chemistry and computermodeling, are used to adjust or modify the structure of the ligand sothat it can make additional or different interactions with the target.

[0182] The ligand design uses computer modeling programs which calculatehow different molecules interact with the various sites of a target.This procedure determines potential ligands or mimetics of theligand(s).

[0183] The ligand design uses computer modeling programs which calculatehow different molecules interact with the various sites of the target,subunit, or a fragment thereof. Thus, this procedure determinespotential ligands or ligand mimetics.

[0184] 6. The newly designed ligand from Step 5 can be tested for itsbiological activity using appropriate in vivo or in vitro tests,including the high throughput screening methods discussed above.

[0185] The potential ligands or mimetics are then screened for activityrelating to HDM2, or at least a fragment thereof. Such screening methodsare selected from assays for at least one biological activity of thenative target.

[0186] The resulting ligands or mimetics, provided by methods of thepresent invention, are useful for treating, screening or preventingdiseases in animals, such as mammals (including humans) and birds.

[0187] 7. Of course, each of the above steps can be modified as desiredby those of skill in the art so as to refine the procedure for theparticular goal in mind. Also, additional X-ray diffraction data may becollected on HDM2, HDM2/ligand complexes, HDM2 structural target motifsand HDM2 subunit/ligand complexes at any step or phase of the procedure.Such additional diffraction data can be used to reconstruct electrondensity maps and molecular models which may further assist in the designand selection of ligands with the desirable binding attributes.

[0188] It is to be understood that the present invention is consideredto include stereoisomers as well as optical isomers, e.g., mixtures ofenantiomers as well as individual enantiomers and diastereomers, whicharise as a consequence of structural asymmetry in selected compounds,ligands or mimetics of the present series.

[0189] Some of the compounds or agents disclosed or discovered by themethods herein may contain one or more asymmetric centers and thus giverise to enantiomers, diastereomers, and other stereoisomeric forms. Thepresent invention is also meant to encompass all such possible forms aswell as their racemic and resolved forms and mixtures thereof. When thecompounds described or discovered herein contain olefinic double bondsor other centers of geometric asymmetry, and unless otherwise specified,it is intended to include both E and Z geometric isomers. All tautomersare intended to be encompassed by the present invention as well.

[0190] As used herein, the term “stereoisomers” is a general term forall isomers of individual molecules that differ only in the orientationof their atoms in space. It includes enantiomers and isomers ofcompounds with more than one chiral center that are not mirror images ofone another (diastereomers).

[0191] As used herein, the term “chiral center” refers to to a carbonatom to which four different groups are attached.

[0192] As used herein, the term “enantiomer” or “enantiomeric” refers toa molecule that is nonsuperimposable on its mirror image and henceoptically active wherein the enantiomer rotates the plane of polarizedlight in one direction and its mirror image rotates the plane ofpolarized light in the opposite direction.

[0193] As used herein, the term “racemic” refers to a mixture of equalparts of enantiomers and which is optically active.

[0194] As used herein, the term “resolution” refers to the separation orconcentration or depletion of one of the two enantiomeric forms of amolecule. In the context of this application. the term “resolution” alsorefers to the amount of detail which can be resolved by the diffractionexperiment. Or in other terms, since the inherent disorder of a proteincrystal diffraction pattern fades away at some diffraction angleθ_(max), the corresponding distance d_(min) of the reciprocal latticesis deterimined by Bragg's law.$d_{\min} = \frac{\lambda}{2\sin \quad \theta_{\max}}$

[0195] In practice in protein crystallography it is usual to quote thenominal resolution of a protein electron density in terms of d_(min),the minimum lattice distance to which data is included in thecalculation of the map.

[0196] The compounds of the present invention are also useful atinhibiting the interaction between p53 and MDMX. MDMX, also known asMDM4, is a cellular protein involved in the regulation of the cellcycle. For example, see Riemenschneider et al., Cancer Res.59(24):6091-6096 (1999).

[0197] Without further description, it is believed that one of ordinaryskill in the art can, using the preceding description and the followingillustrative examples, make and utilize the compounds of the presentinvention and practice the claimed methods. The following workingexamples therefore, specifically point out preferred embodiments of thepresent invention, and are not to be construed as limiting in any waythe remainder of the disclosure.

EXAMPLES Example 1

[0198]

[0199] GST HDM2 Fusion Protein Construction and Expression

[0200] cDNA encoding residues 17-125 of HDM2 were cloned and expressedas follows: PCR was performed using ATCC item number 384988 containingpartial human MDM2 sequence as template and the following primers:Forward: 5′-CTCTCTCGGATCCCAGATTCCAGCTTCGGAACAAGAG Reverse:5′-TATATATCTCGAGTCAGTTCTCACTCACAGATGTACCTGAG.

[0201] The PCR product was then digested with BamHI and Xhol (sequencerecognition sites underlined in primers), gel purified, and ligated intopGEX4t-3 which had also been digested with BamHI and XhoI. The purifiedplasmid was transformed into E. coli strain BL21. Protein was producedat 37° C. in 2 L shake flasks containing 800 ml LB (Laura Bertanimedium)+100 ∥g/ml ampicillin and supplemented with 0.2% glycerol.Briefly media was inoculated with 16 ml of overnight culture and inducedwith 1 mM IPTG when the absorbance at 600 reached 0.6-0.8 OD. Cells wereharvested 5 hr post induction.

[0202] For HDM2 23-114, the primers used were as follows:5′-CGACGATTGGATCCGAACAAAGACCCTG 3′-GGCTACTACTCCGAGTCATTCCTGCTGATTGACTAC

[0203] For HDM2 17-111, the primers used were5′-CTCTCTCGGATCCCAGATTCAGCTTCCGGAACAAGAG3′-TTCAGCAGCTCGAGTCAATTGACTACTACCAAGTTC

[0204] The PCR fragments were cloned and expressed as above with a fewexceptions. E. coli strain BL21 RIL was used for expression. Cells weregrown at 37° C. until A₆₀₀ of 0.2, then transferred to room temperatureand induced at A₆₀₀ of 0.6-0.8 with 0.1 mM IPTG. Cells were harvested 5hours post induction, centrifuged, and resuspended in PBS to 10 ml/gcell paste.

[0205] Protein Production

[0206] Cells were lysed in an Avestin microfluidizer, centrifuged, andthe supernatant bound to a glutathione sepharose 4B resin (Pharmacia).The resin was washed with PBS and the HDM2 construct of interest wascleaved from the GST-resin by the addition of 2 μg/ml thrombin (EnzymeResearch Labs). The cleaved HDM2 was loaded onto a Sepharose SP FastFlow resin (Pharmacia), and eluted with a 20 mM HEPES pH. 7.5, 150 mMNaCl. Glutathione was added to 5 mM, and the protein stored at −70° C.The resulting protein has an N-terminal Glycine before amino acid 17(Serine).

[0207] Protein Preparation for Crystallography

[0208] HDM2 17-111 was complexed with the compound of interest bydialysis at a concentration of 0.7 mg/ml, the buffer brought to 20 mMHEPES pH. 7.4, 100 mM NaCl, 5 mM DTT, filtered through a 0.02 μm filter,and concentrated to 10 mg/ml.

Example 2

[0209] Crystallization and Data Collection

[0210] In a typical crystallization experiment, 1-2 μl of HDM2 protein,complexed with a compound and concentrated to ca. 10 mg/ml, was mixed ina 1:1 ratio with well solution (1.8-2.4M (NH4)₂SO₄, 100 mM buffer pH.6.5-9.0, 2% PEG 400, 100 mM NaSCN) and placed on a glass cover slip. Thecover slip was inverted and sealed over a reservoir of 500-1000 μl ofwell solution and incubated at 4° C. Crystals usually appeared overnight and were ready to harvest after 3-7 days. Crystals were harvestedwith a nylon loop, placed for less than 30 seconds in cryo-solution(2.2M (NH₄)₂SO₄, 100 mM bis-tris-propane pH. 7.5, 2% PEG 400, 100 mMNaSCN, 15% glycerol) and frozen by immersion in liquid nitrogen orliquid propane. Data were collected at 120K on a Bruker AXS M06XCErotating anode and a SMART 6000 CCD detector. The diffraction data wasprocessed with the Proteum suite (Bruker AXS).

Example 3

[0211] Assay Methods: Peptide Binding Assay

[0212] The inhibition of MDM2 binding to p53 was measured using a p53peptide analog binding to MDM2 residues 17-125. The published crystalstructure of this complex (Kussie, P. H., et al., Science 274:948-953(1996)) validates this fragment as containing the p53 binding site, andwe have solved the X-ray structure of the p53 peptide analogMPRFMDYWEGLN, described to be a peptide inhibitor of the MDM2 p53interaction (Böttger, A., et al., J Mol Biol 269:744-756 (1997)). Theassay uses N terminal fluorescein RFMDYWEGL peptide (F1 9 mer). Compoundwas incubated for 15 minutes with 30 nM fluorescein peptide F1 9 mer and120 nM HDM2 17-125 in 50 mM HEPES pH. 7.5, 150 mM NaCl, 3 mM octylglucoside. The polarization of the fluorescein label was measured byexcitation at 485 nm and emission at 530 nm. Polarization was expressedas a percent of a no compound control, using no MDM2 with F1 9 mer asbackground.

Example 4

[0213] HDM2 Atomic Coordinates: Table 1 (Compound 1)

[0214] Table 1 describes the 3-dimensional atomic coordinates of HDM2complexed with compound 1 (338437)((4-Chloro-phenyl)-[3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-aceticacid and bound waters in standard pdb-format. The relevantcrystallographic data are contained in the REMARK section of Table 1.Two molecules of HDM2, related by non-crystallographic symmetry, arepresent in the asymmetric unit and are identified by the CHAINID of Afor the first molecule and B for the second molecule. The compound(compound 1) is present under the residue name DCB. Compound 1 and HDM2molecule sharing the same CHAINID are forming a complex.

Example 5

[0215] HDM2 Atomic Coordinates: Table 2 (Compound 2)

[0216] Compound 2[8-Chloro-3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-(4-chloro-phenyl)-aceticacid (876273) and HDM2 protein were cocrystallized as described above inExample 2. Table 2 describes the 3-dimensional atomic coordinates ofHDM2 complexed with compound 2 (876273)([8-Chloro-3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-(4-chloro-phenyl)-aceticacid). The relevant crystallographic data are contained in the REMARKsection of Table 2. Data were collected as described above. Differentcrystal forms can be observed under the same crystallization conditionsused to obtain the trigonal crystal form.

Example 6

[0217] HDM2 Atomic Coordinates: Table 3

[0218] Table 3 describes the 3-dimensional atomic coordinates of HDM2cocrystallized with compound 2 (compound 876273:[8-Chloro-3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-(4-chloro-phenyl)-aceticacid) in tetragonal spacegroup aligned to the structure of HDM2complexed with compound 1 (compound 338437((4-Chloro-phenyl)-[3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-aceticacid). The relevant crystallographic data are contained in the REMARKsection of Table 3. Data were collected as described above.

[0219] The pdb-format is described on various sites on the web.Depending on the program crystallographic application minormodifications to this format may be found. A good primer is provided bythis link at CCP4 “www.ccp4.ac.uk/html/pdbformat.html”. A more extendeddescription can be found at the RCSB home page.

Example 7

[0220] Phasing: Model Building and Refinement

[0221] Phases were obtained by molecular replacement using the publishedHDM2-structure as a search model in CNX (Brunger, A. T., et al., P. D.Acta Cryst D54:905-921 (1998); Accelrys Inc.). Alternating cycles ofstructure refinement and model building were carried out according tostandard protocols using CNX and O (Jones, T. A., et al., Acta CrystA47: 110-119 (1991)).

Example 8

[0222] Structural Features of HDM2

[0223]FIG. 1. Ribbon representation of HDM2 bound to compound 1(compound 338437:((4-Chloro-phenyl)-[3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-aceticacid).

[0224]FIG. 2. Fit of compound 1 (compound 338437:((4-Chloro-phenyl)-[3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-aceticacid into the active site of HDM2 presented as a molecular surface.

[0225] Although the present invention has been described in detail withreference to examples above, it is understood that various modificationscan be made without departing from the spirit of the invention. Allcited patents, patent applications and publications and other documentscited in this application are herein incorporated by reference in theirentirety. TABLE 1 compound 338437((4-Chloro-phenyl)-[3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl)-aceticacid REMARK coordinates from restrained individual B-factor refinementREMARK refinement resolution: 500.0-2.6 A REMARK starting r = 0.2398free_r = 0.2763 REMARK final r = 0.2390 free_r = 0.2765 REMARK B rmsdfor bonded mainchain atoms = 1.358 target = 1.5 REMARK B rmsd for bondedsidechain atoms = 1.887 target = 2.0 REMARK B rmsd for angle mainchainatoms = 2.371 target = 2.0 REMARK B rmsd for angle sidechain atoms =2.965 target = 2.5 REMARK rweight = 0.1000 (with wa = 2.71183) REMARKtarget = mlf steps = 30 REMARK sg = P3 (2) 21 a = 98.486 b = 98.486 c =74.038 alpha = 90 beta = 90 gamma = 120 REMARK parameter file 1:MSI_CNX_TOPPAR:protein_rep.param REMARK parameter file 2: dcb.par REMARKparameter file 3: MSI_CNX_TOPPAR:water_rep.param REMARK molecularstructure file: cycle8.psf REMARK input coordinates: minimize.pdb REMARKreflection file = ../M338437_P3221.cv REMARK ncs = none REMARKB-correction resolution: 6.0-2.6 REMARK initial B-factor correctionapplied to fobs: REMARK B11 = 5.509 B22 = 5.509 B33 = −11.019 REMARK B12= 0.263 B13 = 0.000 B23 = 0.000 REMARK B-factor correction applied tocoordinate array B: 0.036 REMARK bulk solvent: (Mask) density level =0.372649 e/A{circumflex over ( )}3, B-factor = 25.2844 A{circumflex over( )}2 REMARK reflections with |Fobs|/sigma_F < 0.0 rejected REMARKreflections with |Fobs| > 10000 * rms(Fobs) rejected REMARK theoreticaltotal number of refl. in resol. range: 13090 (100.0%) REMARK number ofunobserved reflections (no entry or |F| = 0): 176 (1.3%) REMARK numberof reflections rejected: 0 (0.0%) REMARK total number of reflectionsused: 12914 (98.7%) REMARK number of reflections in working set: 11964(91.4%) REMARK number of reflections in test set: 950 (7.3%) CRYST198.486 98.486 74.038 90.00 90.00 120.00 P 32 2 1 REMARK FILENAME =“bindividual.pdb” REMARK Written by CNX VERSION: 2000.12 ATOM 1 C GLY A16 47.235 17.293 23.953 1.00 68.07 A C ATOM 2 O GLY A 16 48.284 16.64623.907 1.00 68.13 A O ATOM 3 N GLY A 16 44.698 17.056 23.726 1.00 66.75A N ATOM 4 CA GLY A 16 46.042 16.904 23.083 1.00 67.77 A C ATOM 5 N SERA 17 47.083 18.352 24.744 1.00 67.81 A N ATOM 6 CA SER A 17 48.15418.831 25.618 1.00 66.82 A C ATOM 7 CB SER A 17 48.407 17.831 26.7431.00 67.50 A C ATOM 8 OG SER A 17 47.247 17.658 27.540 1.00 67.94 A OATOM 9 C SER A 17 49.456 19.118 24.864 1.00 65.58 A C ATOM 10 O SER A 1749.699 20.265 24.476 1.00 66.26 A O ATOM 11 N GLN A 18 50.304 18.10824.657 1.00 63.21 A N ATOM 12 CA GLN A 18 51.543 18.367 23.921 1.0060.51 A C ATOM 13 CB GLN A 18 52.778 17.995 24.735 1.00 60.31 A C ATOM14 CG GLN A 18 53.833 19.070 24.574 1.00 59.93 A C ATOM 15 CD GLN A 1853.200 20.457 24.506 1.00 59.70 A C ATOM 16 OE1 GLN A 18 52.584 20.91925.464 1.00 59.78 A O ATOM 17 NE2 GLN A 18 53.333 21.112 23.362 1.0059.29 A N ATOM 18 C GLN A 18 51.619 17.752 22.529 1.00 58.18 A C ATOM 19O GLN A 18 52.569 17.049 22.158 1.00 57.08 A O ATOM 20 N ILE A 19 50.57318.064 21.776 1.00 54.61 A N ATOM 21 CA ILE A 19 50.380 17.673 20.3991.00 49.59 A C ATOM 22 CB ILE A 19 49.130 16.771 20.256 1.00 47.27 A CATOM 23 CG2 ILE A 19 48.730 16.643 18.802 1.00 46.21 A C ATOM 24 CG1 ILEA 19 49.407 15.403 20.880 1.00 44.27 A C ATOM 25 CD1 ILE A 19 50.56514.675 20.263 1.00 40.62 A C ATOM 26 C ILE A 19 50.112 19.056 19.8061.00 48.49 A C ATOM 27 O ILE A 19 49.344 19.838 20.374 1.00 46.95 A OATOM 28 N PRO A 20 50.765 19.396 18.686 1.00 47.71 A N ATOM 29 CD PRO A20 51.681 18.610 17.840 1.00 46.47 A C ATOM 30 CA PRO A 20 50.521 20.72118.107 1.00 46.58 A C ATOM 31 CB PRO A 20 51.072 20.574 16.695 1.0046.92 A C ATOM 32 CG PRO A 20 52.256 19.667 16.919 1.00 46.74 A C ATOM33 C PRO A 20 49.041 21.112 18.134 1.00 45.69 A C ATOM 34 O PRO A 2048.181 20.376 17.636 1.00 45.19 A O ATOM 35 N ALA A 21 48.751 22.26418.738 1.00 43.36 A N ATOM 36 CA ALA A 21 47.379 22.755 18.832 1.0040.68 A C ATOM 37 CB ALA A 21 47.371 24.193 19.350 1.00 39.15 A C ATOM38 C ALA A 21 46.710 22.676 17.460 1.00 39.20 A C ATOM 39 O ALA A 2145.518 22.379 17.351 1.00 38.64 A O ATOM 40 N SER A 22 47.490 22.93716.414 1.00 37.13 A N ATOM 41 CA SER A 22 46.996 22.881 15.042 1.0034.91 A C ATOM 42 CB SER A 22 48.140 23.167 14.077 1.00 36.91 A C ATOM43 OG SER A 22 49.177 22.202 14.208 1.00 39.98 A O ATOM 44 C SER A 2246.428 21.494 14.755 1.00 32.16 A C ATOM 45 O SER A 22 45.349 21.35614.179 1.00 32.41 A O ATOM 46 N GLU A 23 47.179 20.474 15.159 1.00 28.39A N ATOM 47 CA GLU A 23 46.785 19.084 14.981 1.00 25.53 A C ATOM 48 CBGLU A 23 47.986 18.167 15.280 1.00 24.74 A C ATOM 49 CG GLU A 23 47.65016.698 15.507 1.00 22.09 A C ATOM 50 CD GLU A 23 48.881 15.805 15.5241.00 21.23 A C ATOM 51 OE1 GLU A 23 49.956 16.270 15.952 1.00 22.29 A OATOM 52 OE2 GLU A 23 48.775 14.631 15.120 1.00 19.31 A O ATOM 53 C GLU A23 45.597 18.733 15.879 1.00 23.76 A C ATOM 54 O GLU A 23 44.756 17.92815.501 1.00 21.78 A O ATOM 55 N GLN A 24 45.524 19.345 17.059 1.00 22.33A N ATOM 56 CA GLN A 24 44.423 19.086 17.985 1.00 21.86 A C ATOM 57 CBGLN A 24 44.628 19.838 19.294 1.00 22.56 A C ATOM 58 CG GLN A 24 45.72119.267 20.157 1.00 26.35 A C ATOM 59 CD GLN A 24 45.896 20.017 21.4581.00 27.27 A C ATOM 60 OE1 GLN A 24 44.964 20.131 22.262 1.00 27.74 A OATOM 61 NE2 GLN A 24 47.101 20.533 21.676 1.00 29.00 A N ATOM 62 C GLN A24 43.063 19.464 17.423 1.00 21.99 A C ATOM 63 O GLN A 24 42.047 18.87117.786 1.00 21.50 A O ATOM 64 N GLU A 25 43.045 20.456 16.542 1.00 22.95A N ATOM 65 CA GLU A 25 41.800 20.921 15.939 1.00 24.26 A C ATOM 66 CBGLU A 25 41.874 22.432 15.665 1.00 25.71 A C ATOM 67 CG GLU A 25 42.22623.265 16.884 1.00 27.74 A C ATOM 68 CD GLU A 25 41.218 23.107 18.0061.00 30.63 A C ATOM 69 OE1 GLU A 25 41.640 23.112 19.187 1.00 32.01 A OATOM 70 OE2 GLU A 25 40.005 22.987 17.705 1.00 30.20 A O ATOM 71 C GLU A25 41.479 20.179 14.644 1.00 23.62 A C ATOM 72 O GLU A 25 40.472 20.46113.995 1.00 24.16 A O ATOM 73 N THR A 26 42.337 19.237 14.267 1.00 22.53A N ATOM 74 CA THR A 26 42.123 18.462 13.052 1.00 21.97 A C ATOM 75 CBTHR A 26 43.138 17.299 12.955 1.00 23.07 A C ATOM 76 OG1 THR A 26 44.47217.828 12.972 1.00 22.96 A O ATOM 77 CG2 THR A 26 42.920 16.499 11.6751.00 21.12 A C ATOM 78 C THR A 26 40.705 17.894 13.031 1.00 21.66 A CATOM 79 O THR A 26 40.281 17.217 13.962 1.00 19.94 A O ATOM 80 N LEU A27 39.974 18.187 11.963 1.00 23.11 A N ATOM 81 CA LEU A 27 38.602 17.71311.805 1.00 25.25 A C ATOM 82 CB LEU A 27 37.888 18.593 10.775 1.0026.19 A C ATOM 83 CG LEU A 27 36.362 18.646 10.828 1.00 28.80 A C ATOM84 CD1 LEU A 27 35.918 19.212 12.183 1.00 26.94 A C ATOM 85 CD2 LEU A 2735.840 19.512 9.677 1.00 28.47 A C ATOM 86 C LEU A 27 38.620 16.24011.350 1.00 24.95 A C ATOM 87 O LEU A 27 39.275 15.901 10.359 1.00 26.23A O ATOM 88 N VAL A 28 37.898 15.373 12.064 1.00 22.99 A N ATOM 89 CAVAL A 28 37.882 13.938 11.749 1.00 20.72 A C ATOM 90 CB VAL A 28 38.81013.143 12.719 1.00 18.95 A C ATOM 91 CG1 VAL A 28 40.213 13.729 12.7341.00 17.23 A C ATOM 92 CG2 VAL A 28 38.230 13.164 14.118 1.00 16.75 A CATOM 93 C VAL A 28 36.500 13.284 11.833 1.00 21.21 A C ATOM 94 O VAL A28 35.593 13.805 12.486 1.00 19.81 A O ATOM 95 N ARG A 29 36.365 12.13111.174 1.00 22.25 A N ATOM 96 CA ARG A 29 35.126 11.340 11.159 1.0023.75 A C ATOM 97 CB ARG A 29 34.559 11.229 9.742 1.00 25.97 A C ATOM 98CG ARG A 29 33.678 12.388 9.308 1.00 32.69 A C ATOM 99 CD ARG A 2933.206 12.199 7.866 1.00 38.68 A C ATOM 100 NE ARG A 29 32.256 13.2317.455 1.00 45.71 A N ATOM 101 CZ ARG A 29 31.002 13.322 7.901 1.00 50.53A C ATOM 102 NH1 ARG A 29 30.534 12.439 8.777 1.00 52.47 A N ATOM 103NH2 ARG A 29 30.208 14.299 7.472 1.00 52.60 A N ATOM 104 C ARG A 2935.402 9.931 11.680 1.00 22.61 A C ATOM 105 O ARG A 29 35.891 9.07610.944 1.00 23.74 A O ATOM 106 N PRO A 30 35.094 9.669 12.959 1.00 20.77A N ATOM 107 CD PRO A 30 34.654 10.607 14.006 1.00 19.85 A C ATOM 108 CAPRO A 30 35.334 8.340 13.523 1.00 19.49 A C ATOM 109 CB PRO A 30 34.7988.471 14.951 1.00 18.27 A C ATOM 110 CG PRO A 30 35.081 9.895 15.2761.00 18.94 A C ATOM 111 C PRO A 30 34.655 7.206 12.757 1.00 19.01 A CATOM 112 O PRO A 30 33.552 7.367 12.236 1.00 18.19 A O ATOM 113 N LYS A31 35.332 6.063 12.689 1.00 19.05 A N ATOM 114 CA LYS A 31 34.790 4.88012.033 1.00 19.18 A C ATOM 115 CB LYS A 31 35.919 3.891 11.703 1.0018.74 A C ATOM 116 CG LYS A 31 36.881 4.392 10.630 1.00 18.49 A C ATOM117 CD LYS A 31 38.048 3.442 10.409 1.00 16.87 A C ATOM 118 CE LYS A 3139.007 3.982 9.359 1.00 16.16 A C ATOM 119 NZ LYS A 31 40.238 3.1619.254 1.00 17.03 A N ATOM 120 C LYS A 31 33.777 4.260 13.009 1.00 19.76A C ATOM 121 O LYS A 31 33.862 4.465 14.221 1.00 19.24 A O ATOM 122 NPRO A 32 32.816 3.480 12.492 1.00 20.78 A N ATOM 123 CD PRO A 32 32.7722.971 11.106 1.00 20.08 A C ATOM 124 CA PRO A 32 31.778 2.840 13.3111.00 20.65 A C ATOM 125 CB PRO A 32 31.376 1.640 12.465 1.00 18.59 A CATOM 126 CG PRO A 32 31.460 2.197 11.079 1.00 19.84 A C ATOM 127 C PRO A32 32.108 2.460 14.761 1.00 21.61 A C ATOM 128 O PRO A 32 31.412 2.87715.695 1.00 20.03 A O ATOM 129 N LEU A 33 33.164 1.679 14.955 1.00 22.55A N ATOM 130 CA LEU A 33 33.522 1.237 16.296 1.00 23.06 A C ATOM 131 CBLEU A 33 34.677 0.236 16.223 1.00 23.79 A C ATOM 132 CG LEU A 33 34.537−0.992 17.135 1.00 24.29 A C ATOM 133 CD1 LEU A 33 33.136 −1.597 17.0201.00 22.52 A C ATOM 134 CD2 LEU A 33 35.597 −2.015 16.747 1.00 23.61 A CATOM 135 C LEU A 33 33.850 2.370 17.260 1.00 23.64 A C ATOM 136 O LEU A33 33.348 2.385 18.382 1.00 25.22 A O ATOM 137 N LEU A 34 34.684 3.31916.838 1.00 23.73 A N ATOM 138 CA LEU A 34 35.033 4.445 17.707 1.0023.80 A C ATOM 139 CB LEU A 34 36.173 5.281 17.108 1.00 22.45 A C ATOM140 CG LEU A 34 36.459 6.603 17.842 1.00 21.72 A C ATOM 141 CD1 LEU A 3436.722 6.324 19.310 1.00 20.49 A C ATOM 142 CD2 LEU A 34 37.647 7.32417.209 1.00 20.12 A C ATOM 143 C LEU A 34 33.829 5.351 17.949 1.00 24.92A C ATOM 144 O LEU A 34 33.641 5.867 19.058 1.00 25.03 A O ATOM 145 NLEU A 35 33.019 5.549 16.911 1.00 24.02 A N ATOM 146 CA LEU A 35 31.8416.394 17.027 1.00 24.15 A C ATOM 147 CB LEU A 35 31.113 6.470 15.6901.00 21.76 A C ATOM 148 CG LEU A 35 30.415 7.787 15.341 1.00 20.69 A CATOM 149 CD1 LEU A 35 29.176 7.489 14.522 1.00 17.66 A C ATOM 150 CD2LEU A 35 30.026 8.526 16.589 1.00 20.82 A C ATOM 151 C LEU A 35 30.8895.849 18.101 1.00 26.94 A C ATOM 152 O LEU A 35 30.302 6.617 18.870 1.0027.88 A O ATOM 153 N LYS A 36 30.735 4.527 18.152 1.00 27.77 A N ATOM154 CA LYS A 36 29.859 3.913 19.140 1.00 28.78 A C ATOM 155 CB LYS A 3629.778 2.398 18.939 1.00 31.12 A C ATOM 156 CG LYS A 36 29.062 1.68720.081 1.00 35.08 A C ATOM 157 CD LYS A 36 29.472 0.220 20.223 1.0038.33 A C ATOM 158 CE LYS A 36 28.711 −0.681 19.268 1.00 41.52 A C ATOM159 NZ LYS A 36 29.044 −2.120 19.487 1.00 43.62 A N ATOM 160 C LYS A 3630.397 4.200 20.530 1.00 28.98 A C ATOM 161 O LYS A 36 29.632 4.46321.459 1.00 29.52 A O ATOM 162 N LEU A 37 31.719 4.137 20.664 1.00 28.82A N ATOM 163 CA LEU A 37 32.396 4.395 21.936 1.00 28.09 A C ATOM 164 CBLEU A 37 33.908 4.244 21.751 1.00 28.11 A C ATOM 165 CG LEU A 37 34.8884.330 22.929 1.00 27.55 A C ATOM 166 CD1 LEU A 37 34.655 5.592 23.7361.00 27.78 A C ATOM 167 CD2 LEU A 37 34.730 3.109 23.788 1.00 27.77 A CATOM 168 C LEU A 37 32.079 5.814 22.402 1.00 28.36 A C ATOM 169 O LEU A37 31.730 6.034 23.559 1.00 27.57 A O ATOM 170 N LEU A 38 32.209 6.77221.486 1.00 28.83 A N ATOM 171 CA LEU A 38 31.951 8.175 21.785 1.0027.42 A C ATOM 172 CB LEU A 38 32.324 9.045 20.581 1.00 25.41 A C ATOM173 CG LEU A 38 33.787 8.973 20.141 1.00 25.18 A C ATOM 174 CD1 LEU A 3834.015 9.980 19.042 1.00 24.55 A C ATOM 175 CD2 LEU A 38 34.715 9.25121.315 1.00 25.40 A C ATOM 176 C LEU A 38 30.504 8.449 22.186 1.00 27.27A C ATOM 177 O LEU A 38 30.239 8.989 23.259 1.00 27.56 A O ATOM 178 NLYS A 39 29.562 8.083 21.327 1.00 26.08 A N ATOM 179 CA LYS A 39 28.1638.325 21.638 1.00 24.78 A C ATOM 180 CB LYS A 39 27.294 7.885 20.4611.00 23.77 A C ATOM 181 CG LYS A 39 27.593 8.696 19.215 1.00 25.97 A CATOM 182 CD LYS A 39 26.657 8.409 18.051 1.00 26.72 A C ATOM 183 CE LYSA 39 26.943 9.401 16.916 1.00 29.24 A C ATOM 184 NZ LYS A 39 26.1699.183 15.657 1.00 31.08 A N ATOM 185 C LYS A 39 27.708 7.655 22.940 1.0023.69 A C ATOM 186 O LYS A 39 26.812 8.156 23.623 1.00 24.40 A O ATOM187 N SER A 40 28.338 6.545 23.305 1.00 21.60 A N ATOM 188 CA SER A 4027.946 5.850 24.522 1.00 18.78 A C ATOM 189 CB SER A 40 28.602 4.46524.593 1.00 16.27 A C ATOM 190 OG SER A 40 29.947 4.538 25.020 1.0013.26 A O ATOM 191 C SER A 40 28.299 6.663 25.766 1.00 18.97 A C ATOM192 O SER A 40 27.808 6.376 26.854 1.00 18.43 A O ATOM 193 N VAL A 4129.157 7.667 25.616 1.00 18.55 A N ATOM 194 CA VAL A 41 29.515 8.49626.758 1.00 19.39 A C ATOM 195 CB VAL A 41 31.057 8.645 26.940 1.0020.54 A C ATOM 196 CG1 VAL A 41 31.630 7.402 27.607 1.00 18.57 A C ATOM197 CG2 VAL A 41 31.733 8.891 25.600 1.00 21.46 A C ATOM 198 C VAL A 4128.887 9.881 26.662 1.00 20.20 A C ATOM 199 O VAL A 41 29.218 10.77127.444 1.00 19.90 A O ATOM 200 N GLY A 42 27.975 10.062 25.707 1.0020.98 A N ATOM 201 CA GLY A 42 27.306 11.345 25.571 1.00 21.00 A C ATOM202 C GLY A 42 27.542 12.155 24.309 1.00 22.06 A C ATOM 203 O GLY A 4226.809 13.107 24.060 1.00 23.06 A O ATOM 204 N ALA A 43 28.557 11.80723.522 1.00 23.02 A N ATOM 205 CA ALA A 43 28.841 12.530 22.280 1.0023.42 A C ATOM 206 CB ALA A 43 30.075 11.940 21.593 1.00 20.28 A C ATOM207 C ALA A 43 27.632 12.427 21.354 1.00 23.40 A C ATOM 208 O ALA A 4326.816 11.517 21.495 1.00 21.98 A O ATOM 209 N GLN A 44 27.505 13.35220.407 1.00 25.42 A N ATOM 210 CA GLN A 44 26.369 13.279 19.502 1.0027.06 A C ATOM 211 CB GLN A 44 25.130 13.872 20.165 1.00 27.82 A C ATOM212 CG GLN A 44 25.315 15.232 20.763 1.00 28.96 A C ATOM 213 CD GLN A 4424.576 15.357 22.085 1.00 32.56 A C ATOM 214 OE1 GLN A 44 24.357 16.46222.585 1.00 34.47 A O ATOM 215 NE2 GLN A 44 24.200 14.215 22.668 1.0031.28 A N ATOM 216 C GLN A 44 26.520 13.854 18.106 1.00 26.84 A C ATOM217 O GLN A 44 25.532 14.268 17.500 1.00 26.83 A O ATOM 218 N LYS A 4527.745 13.881 17.592 1.00 25.93 A N ATOM 219 CA LYS A 45 27.980 14.36316.230 1.00 25.65 A C ATOM 220 CB LYS A 45 28.960 15.544 16.198 1.0025.47 A C ATOM 221 CG LYS A 45 28.546 16.767 16.992 1.00 24.64 A C ATOM222 CD LYS A 45 29.614 17.860 16.948 1.00 21.21 A C ATOM 223 CE LYS A 4530.907 17.410 17.615 1.00 19.16 A C ATOM 224 NZ LYS A 45 31.849 18.54317.785 1.00 16.01 A N ATOM 225 C LYS A 45 28.627 13.199 15.508 1.0024.50 A C ATOM 226 O LYS A 45 28.904 12.162 16.112 1.00 24.58 A O ATOM227 N ASP A 46 28.861 13.359 14.219 1.00 22.87 A N ATOM 228 CA ASP A 4629.539 12.314 13.483 1.00 23.75 A C ATOM 229 CB ASP A 46 28.765 11.93112.219 1.00 26.21 A C ATOM 230 CG ASP A 46 27.451 11.222 12.525 1.0028.44 A C ATOM 231 OD1 ASP A 46 27.379 10.480 13.530 1.00 28.33 A O ATOM232 OD2 ASP A 46 26.489 11.396 11.746 1.00 30.59 A O ATOM 233 C ASP A 4630.913 12.872 13.125 1.00 23.42 A C ATOM 234 O ASP A 46 31.820 12.13412.759 1.00 24.56 A O ATOM 235 N THR A 47 31.062 14.188 13.245 1.0022.36 A N ATOM 236 CA THR A 47 32.326 14.847 12.939 1.00 20.77 A C ATOM237 CB THR A 47 32.151 15.896 11.831 1.00 21.07 A C ATOM 238 OG1 THR A47 31.573 15.276 10.676 1.00 22.41 A O ATOM 239 CG2 THR A 47 33.49616.498 11.458 1.00 19.56 A C ATOM 240 C THR A 47 32.859 15.536 14.1871.00 19.61 A C ATOM 241 O THR A 47 32.114 16.213 14.893 1.00 19.20 A OATOM 242 N TYR A 48 34.147 15.364 14.461 1.00 17.90 A N ATOM 243 CA TYRA 48 34.744 15.966 15.646 1.00 17.29 A C ATOM 244 CB TYR A 48 34.90414.924 16.763 1.00 17.51 A C ATOM 245 CG TYR A 48 33.645 14.195 17.1781.00 18.47 A C ATOM 246 CD1 TYR A 48 33.094 13.199 16.376 1.00 17.60 A CATOM 247 CE1 TYR A 48 31.926 12.549 16.743 1.00 19.18 A C ATOM 248 CD2TYR A 48 32.993 14.518 18.368 1.00 18.07 A C ATOM 249 CE2 TYR A 4831.826 13.877 18.746 1.00 18.21 A C ATOM 250 CZ TYR A 48 31.291 12.89317.930 1.00 19.97 A C ATOM 251 OH TYR A 48 30.112 12.264 18.294 1.0020.10 A O ATOM 252 C TYR A 48 36.123 16.528 15.356 1.00 17.64 A C ATOM253 O TYR A 48 36.650 16.401 14.245 1.00 19.08 A O ATOM 254 N THR A 4936.695 17.172 16.364 1.00 16.11 A N ATOM 255 CA THR A 49 38.057 17.67716.267 1.00 16.21 A C ATOM 256 CB THR A 49 38.219 19.061 16.891 1.0015.01 A C ATOM 257 OG1 THR A 49 38.028 18.969 18.309 1.00 14.67 A O ATOM258 CG2 THR A 49 37.212 20.006 16.316 1.00 14.41 A C ATOM 259 C THR A 4938.766 16.672 17.164 1.00 16.33 A C ATOM 260 O THR A 49 38.142 16.09118.057 1.00 16.33 A O ATOM 261 N MET A 50 40.050 16.448 16.937 1.0016.12 A N ATOM 262 CA MET A 50 40.770 15.500 17.765 1.00 16.31 A C ATOM263 CB MET A 50 42.253 15.515 17.411 1.00 16.08 A C ATOM 264 CG MET A 5042.550 14.804 16.100 1.00 15.76 A C ATOM 265 SD MET A 50 42.007 13.08816.174 1.00 15.40 A S ATOM 266 CE MET A 50 43.383 12.364 17.108 1.0012.28 A C ATOM 267 C MET A 50 40.570 15.820 19.236 1.00 17.20 A C ATOM268 O MET A 50 40.300 14.931 20.034 1.00 18.81 A O ATOM 269 N LYS A 5140.679 17.099 19.581 1.00 16.94 A N ATOM 270 CA LYS A 51 40.515 17.55120.952 1.00 16.59 A C ATOM 271 CB LYS A 51 40.588 19.083 20.993 1.0019.46 A C ATOM 272 CG LYS A 51 40.773 19.662 22.382 1.00 24.41 A C ATOM273 CD LYS A 51 41.177 21.132 22.333 1.00 29.49 A C ATOM 274 CE LYS A 5141.633 21.623 23.711 1.00 30.87 A C ATOM 275 NZ LYS A 51 42.111 23.03923.693 1.00 32.84 A N ATOM 276 C LYS A 51 39.195 17.059 21.555 1.0015.64 A C ATOM 277 O LYS A 51 39.133 16.677 22.729 1.00 14.01 A O ATOM278 N GLU A 52 38.139 17.062 20.750 1.00 15.07 A N ATOM 279 CA GLU A 5236.837 16.613 21.234 1.00 15.62 A C ATOM 280 CB GLU A 52 35.738 17.01720.254 1.00 16.29 A C ATOM 281 CG GLU A 52 35.586 18.520 20.127 1.0018.24 A C ATOM 282 CD GLU A 52 34.649 18.921 19.018 1.00 19.03 A C ATOM283 OE1 GLU A 52 34.764 18.341 17.918 1.00 22.90 A O ATOM 284 OE2 GLU A52 33.812 19.821 19.236 1.00 19.46 A O ATOM 285 C GLU A 52 36.808 15.11021.454 1.00 15.03 A C ATOM 286 O GLU A 52 36.232 14.638 22.435 1.0016.38 A O ATOM 287 N VAL A 53 37.432 14.361 20.546 1.00 14.02 A N ATOM288 CA VAL A 53 37.475 12.908 20.661 1.00 12.12 A C ATOM 289 CB VAL A 5338.216 12.274 19.478 1.00 11.38 A C ATOM 290 CG1 VAL A 53 38.129 10.76919.566 1.00 10.68 A C ATOM 291 CG2 VAL A 53 37.612 12.750 18.171 1.0012.22 A C ATOM 292 C VAL A 53 38.191 12.540 21.955 1.00 12.50 A C ATOM293 O VAL A 53 37.691 11.744 22.761 1.00 12.84 A O ATOM 294 N LEU A 5439.362 13.129 22.158 1.00 9.57 A N ATOM 295 CA LEU A 54 40.109 12.87123.365 1.00 10.63 A C ATOM 296 CB LEU A 54 41.365 13.726 23.405 1.0011.63 A C ATOM 297 CG LEU A 54 42.520 13.131 22.609 1.00 11.73 A C ATOM298 CD1 LEU A 54 43.563 14.192 22.418 1.00 14.32 A C ATOM 299 CD2 LEU A54 43.095 11.922 23.344 1.00 12.23 A C ATOM 300 C LEU A 54 39.260 13.15924.581 1.00 12.08 A C ATOM 301 O LEU A 54 39.280 12.398 25.541 1.0013.29 A O ATOM 302 N PHE A 55 38.508 14.256 24.545 1.00 13.96 A N ATOM303 CA PHE A 55 37.653 14.612 25.675 1.00 14.56 A C ATOM 304 CB PHE A 5536.874 15.901 25.391 1.00 15.14 A C ATOM 305 CG PHE A 55 35.984 16.33026.530 1.00 12.59 A C ATOM 306 CD1 PHE A 55 36.478 17.135 27.551 1.0012.50 A C ATOM 307 CD2 PHE A 55 34.674 15.870 26.614 1.00 11.59 A C ATOM308 CE1 PHE A 55 35.679 17.473 28.643 1.00 12.57 A C ATOM 309 CE2 PHE A55 33.869 16.198 27.696 1.00 10.75 A C ATOM 310 CZ PHE A 55 34.37317.001 28.714 1.00 12.37 A C ATOM 311 C PHE A 55 36.654 13.503 25.9981.00 15.87 A C ATOM 312 O PHE A 55 36.537 13.073 27.149 1.00 14.84 A OATOM 313 N TYR A 56 35.916 13.059 24.985 1.00 15.22 A N ATOM 314 CA TYRA 56 34.928 12.013 25.201 1.00 16.95 A C ATOM 315 CB TYR A 56 34.07011.832 23.952 1.00 14.70 A C ATOM 316 CG TYR A 56 33.084 12.953 23.7371.00 14.95 A C ATOM 317 CD1 TYR A 56 32.015 13.148 24.615 1.00 15.85 A CATOM 318 CE1 TYR A 56 31.086 14.169 24.406 1.00 17.59 A C ATOM 319 CD2TYR A 56 33.205 13.810 22.645 1.00 15.37 A C ATOM 320 CE2 TYR A 5632.290 14.833 22.423 1.00 16.46 A C ATOM 321 CZ TYR A 56 31.231 15.00723.303 1.00 18.11 A C ATOM 322 OH TYR A 56 30.311 16.001 23.059 1.0018.28 A O ATOM 323 C TYR A 56 35.598 10.698 25.580 1.00 17.45 A C ATOM324 O TYR A 56 35.117 9.958 26.440 1.00 16.40 A O ATOM 325 N LEU A 5736.717 10.411 24.931 1.00 18.36 A N ATOM 326 CA LEU A 57 37.449 9.19425.217 1.00 17.30 A C ATOM 327 CB LEU A 57 38.621 9.081 24.245 1.0015.20 A C ATOM 328 CG LEU A 57 38.903 7.706 23.644 1.00 16.42 A C ATOM329 CD1 LEU A 57 37.637 7.037 23.166 1.00 16.54 A C ATOM 330 CD2 LEU A57 39.860 7.885 22.499 1.00 18.63 A C ATOM 331 C LEU A 57 37.915 9.33226.675 1.00 17.78 A C ATOM 332 O LEU A 57 38.074 8.350 27.398 1.00 17.21A O ATOM 333 N GLY A 58 38.099 10.576 27.103 1.00 17.15 A N ATOM 334 CAGLY A 58 38.515 10.832 28.464 1.00 16.26 A C ATOM 335 C GLY A 58 37.40610.511 29.438 1.00 17.52 A C ATOM 336 O GLY A 58 37.656 9.909 30.4821.00 18.49 A O ATOM 337 N GLN A 59 36.179 10.906 29.114 1.00 15.94 A NATOM 338 CA GLN A 59 35.071 10.617 30.010 1.00 17.18 A C ATOM 339 CB GLNA 59 33.761 11.199 29.481 1.00 17.70 A C ATOM 340 CG GLN A 59 33.71312.720 29.500 1.00 18.77 A C ATOM 341 CD GLN A 59 34.064 13.281 30.8591.00 19.30 A C ATOM 342 OE1 GLN A 59 33.435 12.939 31.856 1.00 20.86 A OATOM 343 NE2 GLN A 59 35.078 14.145 30.909 1.00 18.39 A N ATOM 344 C GLNA 59 34.966 9.115 30.109 1.00 17.93 A C ATOM 345 O GLN A 59 34.903 8.55231.199 1.00 17.50 A O ATOM 346 N TYR A 60 34.984 8.473 28.949 1.00 19.35A N ATOM 347 CA TYR A 60 34.906 7.023 28.854 1.00 19.97 A C ATOM 348 CBTYR A 60 35.270 6.580 27.450 1.00 18.67 A C ATOM 349 CG TYR A 60 35.0615.121 27.258 1.00 18.93 A C ATOM 350 CD1 TYR A 60 33.783 4.608 27.1141.00 19.95 A C ATOM 351 CE1 TYR A 60 33.573 3.256 26.945 1.00 22.90 A CATOM 352 CD2 TYR A 60 36.139 4.243 27.235 1.00 21.13 A C ATOM 353 CE2TYR A 60 35.946 2.880 27.068 1.00 22.87 A C ATOM 354 CZ TYR A 60 34.6562.393 26.920 1.00 24.39 A C ATOM 355 OH TYR A 60 34.439 1.051 26.7221.00 26.24 A O ATOM 356 C TYR A 60 35.833 6.309 29.837 1.00 20.15 A CATOM 357 O TYR A 60 35.384 5.527 30.682 1.00 19.42 A O ATOM 358 N ILE A61 37.130 6.564 29.701 1.00 20.01 A N ATOM 359 CA ILE A 61 38.116 5.94930.574 1.00 21.24 A C ATOM 360 CB ILE A 61 39.508 6.556 30.346 1.0019.34 A C ATOM 361 CG2 ILE A 61 40.449 6.169 31.481 1.00 17.74 A C ATOM362 CG1 ILE A 61 40.050 6.096 28.993 1.00 18.22 A C ATOM 363 CD1 ILE A61 41.343 6.774 28.602 1.00 17.99 A C ATOM 364 C ILE A 61 37.717 6.17532.020 1.00 22.76 A C ATOM 365 O ILE A 61 37.731 5.261 32.837 1.00 19.91A O ATOM 366 N MET A 62 37.342 7.414 32.309 1.00 26.52 A N ATOM 367 CAMET A 62 36.951 7.827 33.645 1.00 28.53 A C ATOM 368 CB MET A 62 36.7359.338 33.661 1.00 28.55 A C ATOM 369 CG MET A 62 37.153 9.971 34.9481.00 29.96 A C ATOM 370 SD MET A 62 38.843 9.527 35.301 1.00 32.30 A SATOM 371 CE MET A 62 39.644 11.112 35.168 1.00 34.41 A C ATOM 372 C META 62 35.709 7.120 34.180 1.00 29.87 A C ATOM 373 O MET A 62 35.689 6.69135.336 1.00 31.34 A O ATOM 374 N THR A 63 34.677 6.992 33.350 1.00 30.81A N ATOM 375 CA THR A 63 33.450 6.342 33.792 1.00 31.38 A C ATOM 376 CBTHR A 63 32.265 6.625 32.834 1.00 31.49 A C ATOM 377 OG1 THR A 63 31.5055.427 32.649 1.00 33.06 A O ATOM 378 CG2 THR A 63 32.747 7.120 31.5011.00 32.85 A C ATOM 379 C THR A 63 33.588 4.836 34.002 1.00 31.82 A CATOM 380 O THR A 63 33.045 4.301 34.975 1.00 32.68 A O ATOM 381 N LYS A64 34.305 4.149 33.115 1.00 31.19 A N ATOM 382 CA LYS A 64 34.491 2.70633.279 1.00 30.89 A C ATOM 383 CB LYS A 64 34.719 2.032 31.922 1.0029.02 A C ATOM 384 CG LYS A 64 33.600 2.295 30.931 1.00 29.54 A C ATOM385 CD LYS A 64 33.657 1.380 29.723 1.00 28.26 A C ATOM 386 CE LYS A 6433.117 0.005 30.046 1.00 29.17 A C ATOM 387 NZ LYS A 64 32.990 −0.82928.825 1.00 27.96 A N ATOM 388 C LYS A 64 35.661 2.408 34.221 1.00 31.59A C ATOM 389 O LYS A 64 35.926 1.256 34.559 1.00 31.45 A O ATOM 390 NARG A 65 36.345 3.463 34.653 1.00 33.27 A N ATOM 391 CA ARG A 65 37.4933.352 35.551 1.00 34.34 A C ATOM 392 CB ARG A 65 37.043 2.891 36.9351.00 37.67 A C ATOM 393 CG ARG A 65 36.016 3.800 37.560 1.00 43.08 A CATOM 394 CD ARG A 65 35.585 3.283 38.909 1.00 46.77 A C ATOM 395 NE ARGA 65 34.434 4.025 39.407 1.00 51.51 A N ATOM 396 CZ ARG A 65 33.9273.888 40.628 1.00 53.49 A C ATOM 397 NH1 ARG A 65 34.474 3.032 41.4841.00 54.10 A N ATOM 398 NH2 ARG A 65 32.872 4.608 40.993 1.00 54.47 A NATOM 399 C ARG A 65 38.557 2.398 35.021 1.00 33.22 A C ATOM 400 O ARG A65 39.086 1.576 35.770 1.00 32.38 A O ATOM 401 N LEU A 66 38.863 2.51833.731 1.00 31.04 A N ATOM 402 CA LEU A 66 39.868 1.687 33.089 1.0029.81 A C ATOM 403 CB LEU A 66 39.821 1.870 31.564 1.00 28.29 A C ATOM404 CG LEU A 66 38.557 1.471 30.795 1.00 27.60 A C ATOM 405 CD1 LEU A 6638.809 1.611 29.301 1.00 25.14 A C ATOM 406 CD2 LEU A 66 38.170 0.03531.121 1.00 26.60 A C ATOM 407 C LEU A 66 41.272 2.020 33.597 1.00 30.22A C ATOM 408 O LEU A 66 42.251 1.416 33.162 1.00 30.64 A O ATOM 409 NTYR A 67 41.375 2.980 34.514 1.00 30.84 A N ATOM 410 CA TYR A 67 42.6773.365 35.055 1.00 32.60 A C ATOM 411 CB TYR A 67 42.688 4.846 35.4381.00 33.43 A C ATOM 412 CG TYR A 67 41.642 5.226 36.463 1.00 36.96 A CATOM 413 CD1 TYR A 67 41.809 4.922 37.810 1.00 37.60 A C ATOM 414 CE1TYR A 67 40.833 5.254 38.745 1.00 39.51 A C ATOM 415 CD2 TYR A 67 40.4685.875 36.077 1.00 38.20 A C ATOM 416 CE2 TYR A 67 39.488 6.211 37.0041.00 38.78 A C ATOM 417 CZ TYR A 67 39.675 5.898 38.334 1.00 39.55 A CATOM 418 OH TYR A 67 38.703 6.227 39.254 1.00 42.02 A O ATOM 419 C TYR A67 43.037 2.534 36.270 1.00 33.52 A C ATOM 420 O TYR A 67 42.167 1.96836.929 1.00 33.03 A O ATOM 421 N ASP A 68 44.327 2.459 36.567 1.00 35.06A N ATOM 422 CA ASP A 68 44.765 1.699 37.721 1.00 36.88 A C ATOM 423 CBASP A 68 46.146 1.100 37.476 1.00 37.88 A C ATOM 424 CG ASP A 68 46.6580.340 38.674 1.00 38.95 A C ATOM 425 OD1 ASP A 68 45.863 −0.408 39.2811.00 39.74 A O ATOM 426 OD2 ASP A 68 47.850 0.486 39.006 1.00 39.12 A OATOM 427 C ASP A 68 44.795 2.593 38.952 1.00 38.61 A C ATOM 428 O ASP A68 45.391 3.671 38.939 1.00 36.17 A O ATOM 429 N GLU A 69 44.138 2.13840.013 1.00 41.45 A N ATOM 430 CA GLU A 69 44.068 2.889 41.261 1.0044.18 A C ATOM 431 CB GLU A 69 43.392 2.042 42.345 1.00 47.21 A C ATOM432 CG GLU A 69 41.883 1.953 42.222 1.00 50.20 A C ATOM 433 CD GLU A 6941.238 3.322 42.249 1.00 52.69 A C ATOM 434 OE1 GLU A 69 41.619 4.13243.126 1.00 53.29 A O ATOM 435 OE2 GLU A 69 40.352 3.586 41.403 1.0054.11 A O ATOM 436 C GLU A 69 45.421 3.366 41.774 1.00 44.17 A C ATOM437 O GLU A 69 45.615 4.553 42.023 1.00 43.71 A O ATOM 438 N LYS A 7046.349 2.430 41.933 1.00 44.88 A N ATOM 439 CA LYS A 70 47.679 2.73342.441 1.00 45.87 A C ATOM 440 CB LYS A 70 48.260 1.482 43.113 1.0047.39 A C ATOM 441 CG LYS A 70 48.001 0.194 42.326 1.00 50.82 A C ATOM442 CD LYS A 70 48.491 −1.060 43.052 1.00 52.70 A C ATOM 443 CE LYS A 7048.068 −2.333 42.307 1.00 53.19 A C ATOM 444 NZ LYS A 70 48.468 −3.58643.022 1.00 53.35 A N ATOM 445 C LYS A 70 48.639 3.255 41.376 1.00 45.61A C ATOM 446 O LYS A 70 49.687 3.803 41.703 1.00 46.82 A O ATOM 447 NGLN A 71 48.284 3.079 40.107 1.00 45.07 A N ATOM 448 CA GLN A 71 49.1233.539 38.995 1.00 44.59 A C ATOM 449 CB GLN A 71 49.852 2.352 38.3571.00 45.81 A C ATOM 450 CG GLN A 71 51.048 2.754 37.525 1.00 48.21 A CATOM 451 CD GLN A 71 52.085 3.500 38.343 1.00 49.07 A C ATOM 452 OE1 GLNA 71 53.008 4.103 37.795 1.00 49.90 A O ATOM 453 NE2 GLN A 71 51.9403.458 39.665 1.00 49.28 A N ATOM 454 C GLN A 71 48.199 4.207 37.977 1.0042.82 A C ATOM 455 O GLN A 71 48.023 3.733 36.851 1.00 41.55 A O ATOM456 N GLN A 72 47.628 5.327 38.406 1.00 40.92 A N ATOM 457 CA GLN A 7246.658 6.096 37.639 1.00 38.61 A C ATOM 458 CB GLN A 72 46.144 7.23238.519 1.00 39.65 A C ATOM 459 CG GLN A 72 45.172 6.741 39.576 1.0041.28 A C ATOM 460 CD GLN A 72 44.819 7.799 40.590 1.00 41.51 A C ATOM461 OE1 GLN A 72 44.671 8.976 40.253 1.00 42.50 A O ATOM 462 NE2 GLN A72 44.666 7.385 41.842 1.00 41.01 A N ATOM 463 C GLN A 72 46.948 6.63136.247 1.00 35.53 A C ATOM 464 O GLN A 72 46.015 6.973 35.532 1.00 34.95A O ATOM 465 N HIS A 73 48.210 6.712 35.847 1.00 33.43 A N ATOM 466 CAHIS A 73 48.508 7.216 34.512 1.00 31.21 A C ATOM 467 CB HIS A 73 49.8417.975 34.514 1.00 32.43 A C ATOM 468 CG HIS A 73 51.039 7.111 34.7531.00 35.52 A C ATOM 469 CD2 HIS A 73 51.615 6.683 35.901 1.00 36.10 A CATOM 470 ND1 HIS A 73 51.796 6.588 33.725 1.00 36.24 A N ATOM 471 CE1HIS A 73 52.788 5.876 34.230 1.00 36.90 A C ATOM 472 NE2 HIS A 73 52.7015.918 35.548 1.00 37.46 A N ATOM 473 C HIS A 73 48.518 6.076 33.487 1.0029.68 A C ATOM 474 O HIS A 73 48.681 6.297 32.278 1.00 28.72 A O ATOM475 N ILE A 74 48.315 4.859 33.987 1.00 26.57 A N ATOM 476 CA ILE A 7448.281 3.663 33.152 1.00 25.03 A C ATOM 477 CB ILE A 74 49.167 2.53833.745 1.00 23.93 A C ATOM 478 CG2 ILE A 74 48.928 1.244 33.010 1.0023.04 A C ATOM 479 CG1 ILE A 74 50.643 2.933 33.669 1.00 22.73 A C ATOM480 CD1 ILE A 74 51.140 3.210 32.279 1.00 21.40 A C ATOM 481 C ILE A 7446.856 3.135 33.017 1.00 24.39 A C ATOM 482 O ILE A 74 46.199 2.83334.010 1.00 25.39 A O ATOM 483 N VAL A 75 46.387 3.025 31.782 1.00 23.12A N ATOM 484 CA VAL A 75 45.048 2.527 31.506 1.00 22.39 A C ATOM 485 CBVAL A 75 44.413 3.306 30.319 1.00 21.87 A C ATOM 486 CG1 VAL A 75 43.0162.784 30.024 1.00 19.40 A C ATOM 487 CG2 VAL A 75 44.376 4.777 30.6291.00 21.46 A C ATOM 488 C VAL A 75 45.121 1.044 31.120 1.00 23.43 A CATOM 489 O VAL A 75 46.031 0.631 30.396 1.00 24.66 A O ATOM 490 N TYR A76 44.174 0.246 31.604 1.00 22.50 A N ATOM 491 CA TYR A 76 44.115 −1.17331.256 1.00 22.83 A C ATOM 492 CB TYR A 76 44.106 −2.063 32.508 1.0022.60 A C ATOM 493 CG TYR A 76 45.435 −2.130 33.237 1.00 23.26 A C ATOM494 CD1 TYR A 76 45.728 −1.257 34.284 1.00 23.34 A C ATOM 495 CE1 TYR A76 46.964 −1.291 34.928 1.00 24.06 A C ATOM 496 CD2 TYR A 76 46.414−3.041 32.855 1.00 22.74 A C ATOM 497 CE2 TYR A 76 47.654 −3.083 33.4931.00 23.63 A C ATOM 498 CZ TYR A 76 47.926 −2.207 34.526 1.00 23.89 A CATOM 499 OH TYR A 76 49.159 −2.234 35.150 1.00 23.35 A O ATOM 500 C TYRA 76 42.825 −1.367 30.465 1.00 23.68 A C ATOM 501 O TYR A 76 41.727−1.245 31.003 1.00 24.33 A O ATOM 502 N CYS A 77 42.953 −1.663 29.1811.00 24.40 A N ATOM 503 CA CYS A 77 41.780 −1.826 28.342 1.00 26.23 A CATOM 504 CB CYS A 77 41.778 −0.744 27.258 1.00 24.90 A C ATOM 505 SG CYSA 77 43.313 −0.601 26.325 1.00 17.76 A S ATOM 506 C CYS A 77 41.681−3.193 27.692 1.00 29.22 A C ATOM 507 O CYS A 77 41.001 −3.354 26.6761.00 29.98 A O ATOM 508 N SER A 78 42.337 −4.176 28.300 1.00 32.27 A NATOM 509 CA SER A 78 42.366 −5.538 27.775 1.00 35.19 A C ATOM 510 CB SERA 78 43.016 −6.469 28.798 1.00 36.60 A C ATOM 511 OG SER A 78 43.133−7.782 28.279 1.00 38.65 A O ATOM 512 C SER A 78 41.020 −6.119 27.3401.00 36.61 A C ATOM 513 O SER A 78 40.777 −6.309 26.144 1.00 38.66 A OATOM 514 N ASN A 79 40.145 −6.402 28.297 1.00 36.16 A N ATOM 515 CA ASNA 79 38.849 −6.982 27.961 1.00 37.19 A C ATOM 516 CB ASN A 79 38.340−7.871 29.109 1.00 40.14 A C ATOM 517 CG ASN A 79 39.448 −8.682 29.7691.00 42.98 A C ATOM 518 OD1 ASN A 79 40.255 −8.146 30.538 1.00 44.77 A OATOM 519 ND2 ASN A 79 39.495 −9.978 29.471 1.00 42.57 A N ATOM 520 C ASNA 79 37.805 −5.909 27.669 1.00 36.10 A C ATOM 521 O ASN A 79 36.619−6.105 27.936 1.00 36.61 A O ATOM 522 N ASP A 80 38.227 −4.787 27.1011.00 33.75 A N ATOM 523 CA ASP A 80 37.283 −3.716 26.836 1.00 31.35 A CATOM 524 CB ASP A 80 37.630 −2.513 27.708 1.00 33.87 A C ATOM 525 CG ASPA 80 36.563 −1.448 27.672 1.00 34.54 A C ATOM 526 OD1 ASP A 80 35.832−1.315 28.679 1.00 35.54 A O ATOM 527 OD2 ASP A 80 36.453 −0.758 26.6311.00 33.13 A O ATOM 528 C ASP A 80 37.229 −3.280 25.383 1.00 29.65 A CATOM 529 O ASP A 80 38.182 −3.474 24.634 1.00 30.16 A O ATOM 530 N LEU A81 36.109 −2.675 24.995 1.00 27.43 A N ATOM 531 CA LEU A 81 35.932−2.200 23.629 1.00 26.30 A C ATOM 532 CB LEU A 81 34.605 −1.461 23.4751.00 25.12 A C ATOM 533 CG LEU A 81 34.437 −0.778 22.111 1.00 23.81 A CATOM 534 CD1 LEU A 81 34.714 −1.778 21.013 1.00 23.42 A C ATOM 535 CD2LEU A 81 33.040 −0.213 21.971 1.00 23.74 A C ATOM 536 C LEU A 81 37.064−1.278 23.194 1.00 26.69 A C ATOM 537 O LEU A 81 37.416 −1.235 22.0101.00 27.34 A O ATOM 538 N LEU A 82 37.621 −0.529 24.144 1.00 25.53 A NATOM 539 CA LEU A 82 38.717 0.375 23.827 1.00 24.07 A C ATOM 540 CB LEUA 82 39.087 1.217 25.047 1.00 20.46 A C ATOM 541 CG LEU A 82 40.2362.202 24.829 1.00 18.75 A C ATOM 542 CD1 LEU A 82 39.913 3.166 23.6891.00 17.60 A C ATOM 543 CD2 LEU A 82 40.488 2.955 26.112 1.00 18.82 A CATOM 544 C LEU A 82 39.908 −0.468 23.376 1.00 25.11 A C ATOM 545 O LEU A82 40.569 −0.149 22.381 1.00 26.18 A O ATOM 546 N GLY A 83 40.162 −1.55324.105 1.00 24.69 A N ATOM 547 CA GLY A 83 41.249 −2.451 23.760 1.0024.41 A C ATOM 548 C GLY A 83 41.170 −2.920 22.318 1.00 24.29 A C ATOM549 O GLY A 83 42.196 −3.139 21.681 1.00 23.83 A O ATOM 550 N ASP A 8439.958 −3.076 21.796 1.00 24.91 A N ATOM 551 CA ASP A 84 39.803 −3.50320.410 1.00 27.77 A C ATOM 552 CB ASP A 84 38.386 −3.996 20.149 1.0028.89 A C ATOM 553 CG ASP A 84 38.093 −5.295 20.837 1.00 31.01 A C ATOM554 OD1 ASP A 84 39.042 −6.085 21.039 1.00 32.12 A O ATOM 555 OD2 ASP A84 36.913 −5.532 21.160 1.00 31.66 A O ATOM 556 C ASP A 84 40.119 −2.39219.410 1.00 28.99 A C ATOM 557 O ASP A 84 40.587 −2.664 18.297 1.0029.29 A O ATOM 558 N LEU A 85 39.849 −1.146 19.799 1.00 28.12 A N ATOM559 CA LEU A 85 40.104 −0.009 18.926 1.00 26.68 A C ATOM 560 CB LEU A 8539.385 1.241 19.441 1.00 27.62 A C ATOM 561 CG LEU A 85 37.866 1.15519.582 1.00 29.80 A C ATOM 562 CD1 LEU A 85 37.325 2.478 20.104 1.0028.93 A C ATOM 563 CD2 LEU A 85 37.245 0.807 18.234 1.00 30.79 A C ATOM564 C LEU A 85 41.592 0.268 18.866 1.00 25.94 A C ATOM 565 O LEU A 8542.169 0.400 17.783 1.00 25.60 A O ATOM 566 N PHE A 86 42.208 0.35420.042 1.00 24.28 A N ATOM 567 CA PHE A 86 43.636 0.633 20.143 1.0023.89 A C ATOM 568 CB PHE A 86 43.980 1.113 21.563 1.00 23.07 A C ATOM569 CG PHE A 86 43.601 2.548 21.837 1.00 21.05 A C ATOM 570 CD1 PHE A 8642.858 3.284 20.914 1.00 19.37 A C ATOM 571 CD2 PHE A 86 43.992 3.16523.021 1.00 20.29 A C ATOM 572 CE1 PHE A 86 42.509 4.612 21.167 1.0018.94 A C ATOM 573 CE2 PHE A 86 43.649 4.496 23.285 1.00 20.32 A C ATOM574 CZ PHE A 86 42.903 5.220 22.352 1.00 19.94 A C ATOM 575 C PHE A 8644.498 −0.575 19.781 1.00 22.94 A C ATOM 576 O PHE A 86 45.550 −0.43019.156 1.00 22.42 A O ATOM 577 N GLY A 87 44.049 −1.762 20.178 1.0021.80 A N ATOM 578 CA GLY A 87 44.800 −2.969 19.881 1.00 20.71 A C ATOM579 C GLY A 87 45.882 −3.270 20.904 1.00 18.88 A C ATOM 580 O GLY A 8746.874 −3.940 20.604 1.00 17.62 A O ATOM 581 N VAL A 88 45.703 −2.76922.119 1.00 17.60 A N ATOM 582 CA VAL A 88 46.685 −3.014 23.158 1.0017.15 A C ATOM 583 CB VAL A 88 47.642 −1.815 23.356 1.00 16.00 A C ATOM584 CG1 VAL A 88 48.277 −1.430 22.032 1.00 13.25 A C ATOM 585 CG2 VAL A88 46.901 −0.652 23.975 1.00 15.16 A C ATOM 586 C VAL A 88 45.997 −3.29024.472 1.00 17.59 A C ATOM 587 O VAL A 88 44.841 −2.922 24.667 1.0018.43 A O ATOM 588 N PRO A 89 46.699 −3.966 25.390 1.00 18.50 A N ATOM589 CD PRO A 89 47.976 −4.673 25.177 1.00 17.26 A C ATOM 590 CA PRO A 8946.138 −4.287 26.705 1.00 17.37 A C ATOM 591 CB PRO A 89 46.958 −5.49627.137 1.00 16.69 A C ATOM 592 CG PRO A 89 48.310 −5.170 26.575 1.0017.70 A C ATOM 593 C PRO A 89 46.271 −3.115 27.677 1.00 17.13 A C ATOM594 O PRO A 89 45.549 −3.042 28.669 1.00 18.98 A O ATOM 595 N SER A 9047.196 −2.200 27.397 1.00 16.91 A N ATOM 596 CA SER A 90 47.394 −1.04428.273 1.00 16.57 A C ATOM 597 CB SER A 90 48.002 −1.485 29.612 1.0015.12 A C ATOM 598 OG SER A 90 49.329 −1.956 29.439 1.00 13.05 A O ATOM599 C SER A 90 48.291 0.023 27.653 1.00 16.15 A C ATOM 600 O SER A 9049.090 −0.261 26.764 1.00 17.41 A O

[0226] TABLE 2 compound 876273([8-Chloro-3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-(4-chloro-phenyl)-acetic acid)REMARK coordinates from restrained individual B-factor refinement REMARKrefinement resolution: 25 − 2.6 A REMARK starting r = 0.2563 free_r =0.2787 REMARK final r = 0.2553 free_r = 0.2761 REMARK B rmsd for bondedmainchain atoms = 1.483 target = 1.5 REMARK B rmsd for bonded sidechainatoms = 1.740 target = 2.0 REMARK B rmsd for angle mainchain atoms =2.593 target = 2.0 REMARK B rmsd for angle sidechain atoms = 2.780target = 2.5 REMARK rweight = 0.1000 (with wa = 3.71696) REMARK target =mlf steps = 30 REMARK sg = P4(3)2(1)2 a = 54.3 b = 54.3 c = 83.3 alpha =90 beta = 90 gamma = 90 REMARK parameter file 1:MSI_CNX_TOPPAR:protein_rep.param REMARK parameter file 2: ../cid.parREMARK molecular structure file: recycle.psf REMARK input coordinates:anneal_9.pdb REMARK reflection file = ../M876273_2_P43212.cv REMARK ncs= none REMARK B-correction resolution: 6.0 − 2.6 REMARK initial B-factorcorrection applied to fobs: REMARK B11 = −1.189 B22 = −1.189 B33 = 2.379REMARK B12 = 0.000 B13 = 0.000 B23 = 0.000 REMARK B-factor correctionapplied to coordinate array B: −0.119 REMARK bulk solvent: (Mask)density level = 0.341945 e/A{circumflex over ( )}3, B-factor = 22.3925A{circumflex over ( )}2 REMARK reflections with |Fobs|/sigma_F < 0.0rejected REMARK reflections with |Fobs| > 10000 * rms (Fobs) rejectedREMARK theoretical total number of refl. in resol. range: 4173 (100.0%)REMARK number of unobserved reflections (no entry or |F| = 0): 9 (0.2%)REMARK number of reflections rejected: 0 (0.0%) REMARK total number ofreflections used: 4164 (99.8%) REMARK number of reflections in workingset: 3737 (89.6%) REMARK number of reflections in test set: 427 (10.2%)CRYST1 54.300 54.300 83.300 90.00 90.00 90.00 P 43 21 2 REMARK FILENAME= “bindividual.pdb” REMARK Written by CNX VERSION: 2000.12 ATOM 1 C GLYA 16 50.842 45.566 39.472 1.00 68.15 A C ATOM 2 O GLY A 16 49.884 45.42940.244 1.00 68.22 A O ATOM 3 N GLY A 16 51.272 44.956 37.085 1.00 67.11A N ATOM 4 CA GLY A 16 51.225 44.463 38.498 1.00 67.90 A C ATOM 5 N SERA 17 51.601 46.662 39.435 1.00 67.05 A N ATOM 6 CA SER A 17 51.35847.819 40.296 1.00 64.73 A C ATOM 7 CB SER A 17 52.359 47.851 41.4581.00 65.01 A C ATOM 8 OG SER A 17 52.175 46.743 42.330 1.00 63.84 A OATOM 9 C SER A 17 51.495 49.080 39.449 1.00 62.82 A C ATOM 10 O SER A 1751.039 50.157 39.837 1.00 62.75 A O ATOM 11 N GLN A 18 52.130 48.92538.289 1.00 60.52 A N ATOM 12 CA GLN A 18 52.323 50.023 37.346 1.0057.89 A C ATOM 13 CB GLN A 18 53.377 49.652 36.306 1.00 57.50 A C ATOM14 CG GLN A 18 54.800 49.725 36.791 1.00 57.38 A C ATOM 15 CD GLN A 1855.786 49.390 35.687 1.00 58.16 A C ATOM 16 OE1 GLN A 18 55.675 49.89234.565 1.00 56.84 A O ATOM 17 NE2 GLN A 18 56.761 48.543 36.002 1.0058.44 A N ATOM 18 C GLN A 18 51.013 50.299 36.620 1.00 55.87 A C ATOM 19O GLN A 18 50.763 51.414 36.157 1.00 55.83 A O ATOM 20 N ILE A 19 50.18749.261 36.524 1.00 52.87 A N ATOM 21 CA ILE A 19 48.898 49.337 35.8501.00 50.05 A C ATOM 22 CB ILE A 19 48.721 48.131 34.883 1.00 48.06 A CATOM 23 CG2 ILE A 19 47.404 48.239 34.138 1.00 48.17 A C ATOM 24 CG1 ILEA 19 49.885 48.069 33.889 1.00 45.44 A C ATOM 25 CD1 ILE A 19 49.93949.218 32.921 1.00 43.26 A C ATOM 26 C ILE A 19 47.769 49.319 36.8841.00 49.72 A C ATOM 27 O ILE A 19 47.863 48.631 37.902 1.00 49.03 A OATOM 28 N PRO A 20 46.694 50.095 36.643 1.00 49.57 A N ATOM 29 CD PRO A20 46.609 51.167 35.636 1.00 49.58 A C ATOM 30 CA PRO A 20 45.546 50.16237.553 1.00 49.39 A C ATOM 31 CB PRO A 20 44.693 51.271 36.949 1.0048.67 A C ATOM 32 CG PRO A 20 45.704 52.159 36.318 1.00 48.94 A C ATOM33 C PRO A 20 44.784 48.836 37.628 1.00 49.97 A C ATOM 34 O PRO A 2044.551 48.184 36.606 1.00 50.18 A O ATOM 35 N ALA A 21 44.399 48.44638.840 1.00 49.45 A N ATOM 36 CA ALA A 21 43.660 47.207 39.057 1.0049.81 A C ATOM 37 CB ALA A 21 43.252 47.094 40.528 1.00 49.85 A C ATOM38 C ALA A 21 42.419 47.133 38.160 1.00 49.65 A C ATOM 39 O ALA A 2142.160 46.112 37.517 1.00 49.33 A O ATOM 40 N SER A 22 41.650 48.21738.125 1.00 48.76 A N ATOM 41 CA SER A 22 40.451 48.260 37.302 1.0047.75 A C ATOM 42 CB SER A 22 39.873 49.678 37.296 1.00 47.15 A C ATOM43 OG SER A 22 40.857 50.625 36.915 1.00 48.43 A O ATOM 44 C SER A 2240.792 47.816 35.877 1.00 46.34 A C ATOM 45 O SER A 22 40.029 47.08335.242 1.00 45.89 A O ATOM 46 N GLU A 23 41.947 48.251 35.384 1.00 44.68A N ATOM 47 CA GLU A 23 42.375 47.887 34.040 1.00 43.25 A C ATOM 48 CBGLU A 23 43.526 48.771 33.588 1.00 42.16 A C ATOM 49 CG GLU A 23 43.93948.525 32.164 1.00 40.86 A C ATOM 50 CD GLU A 23 44.747 49.671 31.6121.00 40.82 A C ATOM 51 OE1 GLU A 23 45.613 50.189 32.344 1.00 41.52 A OATOM 52 OE2 GLU A 23 44.523 50.054 30.448 1.00 41.56 A O ATOM 53 C GLU A23 42.790 46.428 33.991 1.00 42.01 A C ATOM 54 O GLU A 23 42.419 45.70133.076 1.00 42.32 A O ATOM 55 N GLN A 24 43.561 45.998 34.977 1.00 41.29A N ATOM 56 CA GLN A 24 43.973 44.610 35.027 1.00 42.07 A C ATOM 57 CBGLN A 24 44.762 44.334 36.314 1.00 41.13 A C ATOM 58 CG GLN A 24 46.20644.813 36.250 1.00 42.57 A C ATOM 59 CD GLN A 24 46.978 44.602 37.5461.00 43.91 A C ATOM 60 OE1 GLN A 24 46.823 43.580 38.225 1.00 44.84 A OATOM 61 NE2 GLN A 24 47.831 45.564 37.884 1.00 43.17 A N ATOM 62 C GLN A24 42.714 43.747 34.984 1.00 43.32 A C ATOM 63 O GLN A 24 42.750 42.59634.541 1.00 43.40 A O ATOM 64 N GLU A 25 41.596 44.326 35.423 1.00 44.62A N ATOM 65 CA GLU A 25 40.314 43.618 35.464 1.00 44.92 A C ATOM 66 CBGLU A 25 39.471 44.123 36.642 1.00 48.33 A C ATOM 67 CG GLU A 25 40.21644.254 37.972 1.00 53.04 A C ATOM 68 CD GLU A 25 40.899 42.969 38.4231.00 55.79 A C ATOM 69 OE1 GLU A 25 41.474 42.972 39.533 1.00 57.11 A OATOM 70 OE2 GLU A 25 40.869 41.961 37.681 1.00 57.84 A O ATOM 71 C GLU A25 39.472 43.697 34.184 1.00 42.75 A C ATOM 72 O GLU A 25 38.563 42.88733.992 1.00 42.62 A O ATOM 73 N THR A 26 39.760 44.666 33.319 1.00 40.18A N ATOM 74 CA THR A 26 39.013 44.813 32.067 1.00 38.13 A C ATOM 75 CBTHR A 26 39.714 45.803 31.100 1.00 37.92 A C ATOM 76 OG1 THR A 26 40.06147.002 31.803 1.00 37.02 A O ATOM 77 CG2 THR A 26 38.798 46.161 29.9451.00 36.76 A C ATOM 78 C THR A 26 38.897 43.456 31.367 1.00 36.79 A CATOM 79 O THR A 26 39.859 42.693 31.321 1.00 36.47 A O ATOM 80 N LEU A27 37.715 43.152 30.841 1.00 36.63 A N ATOM 81 CA LEU A 27 37.490 41.88730.138 1.00 35.77 A C ATOM 82 CB LEU A 27 36.035 41.429 30.291 1.0036.16 A C ATOM 83 CG LEU A 27 35.799 39.909 30.293 1.00 38.05 A C ATOM84 CD1 LEU A 27 36.293 39.313 31.617 1.00 36.12 A C ATOM 85 CD2 LEU A 2734.305 39.610 30.112 1.00 38.41 A C ATOM 86 C LEU A 27 37.808 42.13028.665 1.00 34.52 A C ATOM 87 O LEU A 27 37.395 43.139 28.090 1.00 33.72A O ATOM 88 N VAL A 28 38.540 41.204 28.054 1.00 33.53 A N ATOM 89 CAVAL A 28 38.941 41.364 26.662 1.00 32.88 A C ATOM 90 CB VAL A 28 40.42041.848 26.589 1.00 32.86 A C ATOM 91 CG1 VAL A 28 40.570 43.182 27.3021.00 31.21 A C ATOM 92 CG2 VAL A 28 41.340 40.821 27.242 1.00 33.40 A CATOM 93 C VAL A 28 38.792 40.107 25.802 1.00 31.29 A C ATOM 94 O VAL A28 38.708 38.992 26.314 1.00 30.99 A O ATOM 95 N ARG A 29 38.754 40.31224.489 1.00 29.80 A N ATOM 96 CA ARG A 29 38.641 39.224 23.528 1.0028.94 A C ATOM 97 CB ARG A 29 37.379 39.371 22.685 1.00 33.14 A C ATOM98 CG ARG A 29 36.132 38.818 23.326 1.00 38.54 A C ATOM 99 CD ARG A 2934.905 39.188 22.511 1.00 43.39 A C ATOM 100 NE ARG A 29 33.712 38.54923.051 1.00 47.28 A N ATOM 101 CZ ARG A 29 33.397 37.276 22.843 1.0049.30 A C ATOM 102 NH1 ARG A 29 34.185 36.511 22.093 1.00 49.75 A N ATOM103 nh2 ARG A 29 32.309 36.763 23.405 1.00 50.29 A N ATOM 104 C ARG A 2939.842 39.254 22.602 1.00 26.43 A C ATOM 105 O ARG A 29 39.935 40.12121.727 1.00 25.41 A O ATOM 106 N PRO A 30 40.785 38.315 22.789 1.0024.69 A N ATOM 107 CD PRO A 30 40.798 37.274 23.830 1.00 23.43 A C ATOM108 CA PRO A 30 41.995 38.234 21.958 1.00 23.00 A C ATOM 109 CB PRO A 3042.826 37.151 22.643 1.00 21.72 A C ATOM 110 CG PRO A 30 42.261 37.06424.025 1.00 22.48 A C ATOM 111 C PRO A 30 41.620 37.801 20.544 1.0022.20 A C ATOM 112 O PRO A 30 40.663 37.050 20.360 1.00 22.27 A O ATOM113 N LYS A 31 42.365 38.273 19.551 1.00 21.65 A N ATOM 114 CA LYS A 3142.118 37.880 18.166 1.00 19.51 A C ATOM 115 CB LYS A 31 42.825 38.83917.210 1.00 19.68 A C ATOM 116 CG LYS A 31 42.364 40.279 17.348 1.0020.82 A C ATOM 117 CD LYS A 31 43.115 41.174 16.376 1.00 22.74 A C ATOM118 CE LYS A 31 42.641 42.630 16.435 1.00 20.68 A C ATOM 119 NZ LYS A 3143.356 43.433 15.396 1.00 21.75 A N ATOM 120 C LYS A 31 42.666 36.45418.011 1.00 18.36 A C ATOM 121 O LYS A 31 43.441 35.983 18.847 1.0017.38 A O ATOM 122 N PRO A 32 42.291 35.760 16.930 1.00 17.15 A N ATOM123 CD PRO A 32 41.612 36.293 15.736 1.00 16.30 A C ATOM 124 CA PRO A 3242.737 34.387 16.684 1.00 16.70 A C ATOM 125 CB PRO A 32 42.392 34.18215.213 1.00 16.53 A C ATOM 126 CG PRO A 32 41.180 35.032 15.044 1.0016.35 A C ATOM 127 C PRO A 32 44.198 34.043 16.997 1.00 16.73 A C ATOM128 O PRO A 32 44.470 33.062 17.695 1.00 17.12 A O ATOM 129 N LEU A 3345.137 34.830 16.483 1.00 14.62 A N ATOM 130 CA LEU A 33 46.540 34.53816.724 1.00 13.59 A C ATOM 131 CB LEU A 33 47.425 35.427 15.843 1.0015.36 A C ATOM 132 CG LEU A 33 48.097 34.759 14.626 1.00 13.91 A C ATOM133 CD1 LEU A 33 47.597 33.319 14.406 1.00 13.51 A C ATOM 134 CD2 LEU A33 47.838 35.609 13.411 1.00 8.87 A C ATOM 135 C LEU A 33 46.936 34.64118.189 1.00 13.14 A C ATOM 136 O LEU A 33 47.545 33.724 18.715 1.0012.37 A O ATOM 137 N LEU A 34 46.610 35.744 18.856 1.00 14.58 A N ATOM138 CA LEU A 34 46.930 35.859 20.279 1.00 13.84 A C ATOM 139 CB LEU A 3446.540 37.236 20.826 1.00 13.73 A C ATOM 140 CG LEU A 34 46.613 37.44922.347 1.00 10.65 A C ATOM 141 CD1 LEU A 34 48.041 37.367 22.818 1.009.38 A C ATOM 142 CD2 LEU A 34 46.029 38.795 22.701 1.00 11.47 A C ATOM143 C LEU A 34 46.158 34.771 21.039 1.00 15.75 A C ATOM 144 O LEU A 3446.678 34.176 21.975 1.00 16.50 A O ATOM 145 N LEU A 35 44.917 34.50620.635 1.00 17.07 A N ATOM 146 CA LEU A 35 44.125 33.469 21.295 1.0018.94 A C ATOM 147 CB LEU A 35 42.720 33.381 20.686 1.00 16.70 A C ATOM148 CG LEU A 35 41.753 32.424 21.391 1.00 14.51 A C ATOM 149 CD1 LEU A35 41.542 32.888 22.825 1.00 13.73 A C ATOM 150 CD2 LEU A 35 40.42532.379 20.653 1.00 13.01 A C ATOM 151 C LEU A 35 44.815 32.094 21.2181.00 20.66 A C ATOM 152 O LEU A 35 44.668 31.276 22.133 1.00 21.36 A OATOM 153 N LYS A 36 45.563 31.845 20.140 1.00 20.77 A N ATOM 154 CA LYSA 36 46.279 30.581 19.979 1.00 22.28 A C ATOM 155 CB LYS A 36 46.79230.398 18.547 1.00 23.62 A C ATOM 156 CG LYS A 36 45.782 29.781 17.5861.00 28.23 A C ATOM 157 CD LYS A 36 46.444 28.754 16.647 1.00 30.56 A CATOM 158 CE LYS A 36 47.556 29.366 15.789 1.00 32.43 A C ATOM 159 NZ LYSA 36 48.108 28.414 14.778 1.00 31.16 A N ATOM 160 C LYS A 36 47.46130.504 20.937 1.00 23.86 A C ATOM 161 O LYS A 36 47.716 29.451 21.5371.00 26.24 A O ATOM 162 N LEU A 37 48.196 31.603 21.076 1.00 23.16 A NATOM 163 CA LEU A 37 49.332 31.612 21.994 1.00 24.32 A C ATOM 164 CB LEUA 37 49.920 33.019 22.134 1.00 25.18 A C ATOM 165 CG LEU A 37 50.64733.606 20.934 1.00 27.89 A C ATOM 166 CD1 LEU A 37 51.343 34.901 21.3431.00 28.24 A C ATOM 167 CD2 LEU A 37 51.659 32.597 20.431 1.00 28.07 A CATOM 168 C LEU A 37 48.884 31.144 23.377 1.00 23.05 A C ATOM 169 O LEU A37 49.526 30.304 24.003 1.00 20.34 A O ATOM 170 N LEU A 38 47.770 31.70623.834 1.00 23.36 A N ATOM 171 CA LEU A 38 47.218 31.399 25.139 1.0024.58 A C ATOM 172 CB LEU A 38 46.036 32.322 25.447 1.00 21.20 A C ATOM173 CG LEU A 38 46.216 33.821 25.189 1.00 19.16 A C ATOM 174 CD1 LEU A38 44.954 34.538 25.619 1.00 16.77 A C ATOM 175 CD2 LEU A 38 47.42034.370 25.950 1.00 18.04 A C ATOM 176 C LEU A 38 46.772 29.950 25.2411.00 27.20 A C ATOM 177 O LEU A 38 46.970 29.309 26.273 1.00 29.33 A OATOM 178 N LYS A 39 46.175 29.419 24.182 1.00 28.13 A N ATOM 179 CA LYSA 39 45.720 28.041 24.241 1.00 29.30 A C ATOM 180 CB LYS A 39 44.78227.741 23.071 1.00 28.63 A C ATOM 181 CG LYS A 39 43.532 28.596 23.1231.00 28.78 A C ATOM 182 CD LYS A 39 42.451 28.113 22.184 1.00 27.98 A CATOM 183 CE LYS A 39 41.239 29.021 22.270 1.00 26.89 A C ATOM 184 NZ LYSA 39 40.089 28.444 21.537 1.00 27.11 A N ATOM 185 C LYS A 39 46.88427.062 24.270 1.00 30.33 A C ATOM 186 O LYS A 39 46.787 25.998 24.8831.00 31.74 A O ATOM 187 N SER A 40 47.993 27.428 23.633 1.00 30.27 A NATOM 188 CA SER A 40 49.169 26.559 23.607 1.00 29.62 A C ATOM 189 CB SERA 40 50.280 27.175 22.735 1.00 28.81 A C ATOM 190 OG SER A 40 50.90428.292 23.357 1.00 26.68 A O ATOM 191 C SER A 40 49.701 26.301 25.0201.00 29.48 A C ATOM 192 O SER A 40 50.396 25.316 25.254 1.00 29.59 A OATOM 193 N VAL A 41 49.374 27.186 25.958 1.00 30.35 A N ATOM 194 CA VALA 41 49.827 27.044 27.345 1.00 30.76 A C ATOM 195 CB VAL A 41 50.71728.252 27.799 1.00 30.64 A C ATOM 196 CG1 VAL A 41 52.087 28.184 27.1251.00 29.00 A C ATOM 197 CG2 VAL A 41 50.030 29.574 27.470 1.00 28.32 A CATOM 198 C VAL A 41 48.678 26.891 28.345 1.00 32.02 A C ATOM 199 O VAL A41 48.691 27.500 29.419 1.00 31.05 A O ATOM 200 N GLY A 42 47.675 26.09227.983 1.00 33.77 A N ATOM 201 CA GLY A 42 46.563 25.862 28.889 1.0036.14 A C ATOM 202 C GLY A 42 45.260 26.597 28.641 1.00 38.10 A C ATOM203 O GLY A 42 44.201 25.971 28.653 1.00 40.02 A O ATOM 204 N ALA A 4345.317 27.912 28.432 1.00 38.97 A N ATOM 205 CA ALA A 43 44.107 28.70328.203 1.00 39.16 A C ATOM 206 CB ALA A 43 44.459 30.022 27.518 1.0038.86 A C ATOM 207 C ALA A 43 43.080 27.938 27.375 1.00 39.94 A C ATOM208 O ALA A 43 43.437 27.164 26.485 1.00 40.04 A O ATOM 209 N GLN A 4441.802 28.155 27.675 1.00 41.54 A N ATOM 210 CA GLN A 44 40.733 27.47526.955 1.00 42.78 A C ATOM 211 CB GLN A 44 40.609 26.037 27.466 1.0045.11 A C ATOM 212 CG GLN A 44 40.573 25.913 28.984 1.00 47.61 A C ATOM213 CD GLN A 44 40.868 24.493 29.456 1.00 49.56 A C ATOM 214 OE1 GLN A44 40.913 24.221 30.662 1.00 49.35 A O ATOM 215 NE2 GLN A 44 41.07523.581 28.506 1.00 49.08 A N ATOM 216 C GLN A 44 39.379 28.183 27.0301.00 42.23 A C ATOM 217 O GLN A 44 38.355 27.557 27.322 1.00 42.70 A OATOM 218 N LYS A 45 39.391 29.488 26.763 1.00 40.15 A N ATOM 219 CA LYSA 45 38.184 30.307 26.765 1.00 37.90 A C ATOM 220 CB LYS A 45 38.08531.150 28.034 1.00 39.62 A C ATOM 221 CG LYS A 45 38.193 30.392 29.3411.00 41.18 A C ATOM 222 CD LYS A 45 37.768 31.288 30.502 1.00 42.96 A CATOM 223 CE LYS A 45 38.491 32.629 30.473 1.00 45.64 A C ATOM 224 NZ LYSA 45 38.045 33.556 31.559 1.00 48.08 A N ATOM 225 C LYS A 45 38.29631.254 25.585 1.00 36.61 A C ATOM 226 O LYS A 45 39.258 31.183 24.8221.00 36.82 A O ATOM 227 N ASP A 46 37.323 32.149 25.448 1.00 34.69 A NATOM 228 CA ASP A 46 37.332 33.128 24.368 1.00 33.64 A C ATOM 229 CB ASPA 46 36.015 33.106 23.577 1.00 35.21 A C ATOM 230 CG ASP A 46 35.82631.828 22.778 1.00 36.60 A C ATOM 231 OD1 ASP A 46 36.819 31.326 22.2041.00 35.63 A O ATOM 232 OD2 ASP A 46 34.677 31.337 22.712 1.00 36.79 A OATOM 233 C ASP A 46 37.529 34.526 24.935 1.00 32.32 A C ATOM 234 O ASP A46 38.032 35.419 24.249 1.00 33.06 A O ATOM 235 N THR A 47 37.118 34.71526.186 1.00 30.30 A N ATOM 236 CA THR A 47 37.233 36.012 26.850 1.0027.52 A C ATOM 237 CB THR A 47 35.849 36.568 27.265 1.00 28.37 A C ATOM238 OG1 THR A 47 35.055 35.513 27.822 1.00 28.12 A O ATOM 239 CG2 THR A47 35.130 37.169 26.071 1.00 28.11 A C ATOM 240 C THR A 47 38.097 35.89728.088 1.00 25.47 A C ATOM 241 O THR A 47 38.111 34.862 28.759 1.0024.25 A O ATOM 242 N TYR A 48 38.820 36.973 28.382 1.00 24.41 A N ATOM243 CA TYR A 48 39.716 37.010 29.526 1.00 24.02 A C ATOM 244 CB TYR A 4841.142 36.566 29.126 1.00 23.01 A C ATOM 245 CG TYR A 48 41.265 35.19228.492 1.00 22.29 A C ATOM 246 CD1 TYR A 48 40.966 34.992 27.143 1.0021.16 A C ATOM 247 CE1 TYR A 48 41.035 33.713 26.568 1.00 20.20 A C ATOM248 CD2 TYR A 48 41.647 34.079 29.254 1.00 21.34 A C ATOM 249 CE2 TYR A48 41.721 32.806 28.688 1.00 20.29 A C ATOM 250 CZ TYR A 48 41.40932.633 27.348 1.00 19.58 A C ATOM 251 OH TYR A 48 41.436 31.378 26.7951.00 20.11 A O ATOM 252 C TYR A 48 39.829 38.419 30.085 1.00 24.38 A CATOM 253 O TYR A 48 39.512 39.403 29.409 1.00 22.46 A O ATOM 254 N THR A49 40.288 38.497 31.329 1.00 24.50 A N ATOM 255 CA THR A 49 40.55439.774 31.974 1.00 25.25 A C ATOM 256 CB THR A 49 40.510 39.647 33.5151.00 26.57 A C ATOM 257 OG1 THR A 49 41.215 38.459 33.914 1.00 26.24 A OATOM 258 CG2 THR A 49 39.056 39.580 34.017 1.00 24.91 A C ATOM 259 C THRA 49 42.000 40.027 31.524 1.00 25.63 A C ATOM 260 O THR A 49 42.73839.079 31.248 1.00 24.71 A O ATOM 261 N MET A 50 42.414 41.281 31.4241.00 26.39 A N ATOM 262 CA MET A 50 43.776 41.545 30.986 1.00 26.62 A CATOM 263 CB MET A 50 44.053 43.040 30.977 1.00 25.74 A C ATOM 264 CG META 50 43.446 43.740 29.780 1.00 25.74 A C ATOM 265 SD MET A 50 44.28043.286 28.239 1.00 24.59 A S ATOM 266 CE MET A 50 45.652 44.433 28.2251.00 22.52 A C ATOM 267 C MET A 50 44.797 40.832 31.855 1.00 28.23 A CATOM 268 O MET A 50 45.890 40.518 31.396 1.00 29.67 A O ATOM 269 N LYS A51 44.442 40.554 33.105 1.00 28.66 A N ATOM 270 CA LYS A 51 45.37039.872 33.993 1.00 29.11 A C ATOM 271 CB LYS A 51 44.878 39.943 35.4391.00 32.66 A C ATOM 272 CG LYS A 51 45.919 39.458 36.443 1.00 37.67 A CATOM 273 CD LYS A 51 45.462 39.604 37.889 1.00 41.07 A C ATOM 274 CE LYSA 51 46.527 39.071 38.855 1.00 42.73 A C ATOM 275 NZ LYS A 51 46.10939.129 40.295 1.00 45.66 A N ATOM 276 C LYS A 51 45.597 38.411 33.5861.00 28.29 A C ATOM 277 O LYS A 51 46.723 37.908 33.669 1.00 28.36 A OATOM 278 N GLU A 52 44.537 37.729 33.152 1.00 25.71 A N ATOM 279 CA GLUA 52 44.662 36.334 32.732 1.00 23.60 A C ATOM 280 CB GLU A 52 43.27835.702 32.522 1.00 24.83 A C ATOM 281 CG GLU A 52 42.420 35.589 33.7771.00 27.15 A C ATOM 282 CD GLU A 52 40.989 35.116 33.479 1.00 29.01 A CATOM 283 OE1 GLU A 52 40.281 35.785 32.691 1.00 29.80 A O ATOM 284 OE2GLU A 52 40.568 34.078 34.032 1.00 29.19 A O ATOM 285 C GLU A 52 45.47636.230 31.432 1.00 21.67 A C ATOM 286 O GLU A 52 46.128 35.214 31.1831.00 20.99 A O ATOM 287 N VAL A 53 45.425 37.266 30.598 1.00 18.55 A NATOM 288 CA VAL A 53 46.181 37.255 29.351 1.00 16.46 A C ATOM 289 CB VALA 53 45.807 38.454 28.423 1.00 17.02 A C ATOM 290 CG1 VAL A 53 46.80638.553 27.264 1.00 15.92 A C ATOM 291 CG2 VAL A 53 44.402 38.261 27.8531.00 14.61 A C ATOM 292 C VAL A 53 47.650 37.331 29.742 1.00 15.15 A CATOM 293 O VAL A 53 48.436 36.443 29.403 1.00 13.10 A O ATOM 294 N LEU A54 48.010 38.394 30.463 1.00 15.42 A N ATOM 295 CA LEU A 54 49.37538.570 30.952 1.00 14.74 A C ATOM 296 CB LEU A 54 49.454 39.698 31.9821.00 12.86 A C ATOM 297 CG LEU A 54 49.718 41.124 31.497 1.00 13.00 A CATOM 298 CD1 LEU A 54 50.874 41.100 30.500 1.00 14.60 A C ATOM 299 CD2LEU A 54 48.494 41.706 30.859 1.00 11.73 A C ATOM 300 C LEU A 54 49.84637.281 31.619 1.00 15.13 A C ATOM 301 O LEU A 54 51.009 36.897 31.5021.00 16.88 A O ATOM 302 N PHE A 55 48.942 36.605 32.312 1.00 14.69 A NATOM 303 CA PHE A 55 49.305 35.371 32.991 1.00 16.41 A C ATOM 304 CB PHEA 55 48.137 34.831 33.819 1.00 18.34 A C ATOM 305 CG PHE A 55 48.42833.500 34.429 1.00 20.72 A C ATOM 306 CD1 PHE A 55 49.136 33.411 35.6261.00 20.24 A C ATOM 307 CD2 PHE A 55 48.101 32.324 33.749 1.00 20.52 A CATOM 308 CE1 PHE A 55 49.524 32.170 36.135 1.00 20.60 A C ATOM 309 CE2PHE A 55 48.485 31.079 34.248 1.00 21.73 A C ATOM 310 CZ PHE A 55 49.19931.001 35.444 1.00 20.59 A C ATOM 311 C PHE A 55 49.779 34.268 32.0581.00 16.04 A C ATOM 312 O PHE A 55 50.853 33.698 32.259 1.00 17.31 A OATOM 313 N TYR A 56 48.960 33.949 31.060 1.00 16.71 A N ATOM 314 CA TYRA 56 49.283 32.904 30.087 1.00 17.61 A C ATOM 315 CB TYR A 56 48.07932.620 29.189 1.00 18.45 A C ATOM 316 CG TYR A 56 46.996 31.859 29.9021.00 21.48 A C ATOM 317 CD1 TYR A 56 47.213 30.541 30.318 1.00 23.38 A CATOM 318 CE1 TYR A 56 46.237 29.832 31.013 1.00 25.16 A C ATOM 319 CD2TYR A 56 45.766 32.459 30.198 1.00 22.53 A C ATOM 320 CE2 TYR A 5644.773 31.761 30.898 1.00 23.90 A C ATOM 321 CZ TYR A 56 45.019 30.44531.303 1.00 26.15 A C ATOM 322 OH TYR A 56 44.065 29.738 32.003 1.0026.91 A O ATOM 323 C TYR A 56 50.450 33.345 29.243 1.00 16.78 A C ATOM324 O TYR A 56 51.235 32.530 28.754 1.00 18.53 A O ATOM 325 N LEU A 5750.543 34.656 29.065 1.00 15.78 A N ATOM 326 CA LEU A 57 51.619 35.25228.300 1.00 13.13 A C ATOM 327 CB LEU A 57 51.348 36.749 28.171 1.0011.95 A C ATOM 328 CG LEU A 57 51.232 37.364 26.771 1.00 13.21 A C ATOM329 CD1 LEU A 57 50.535 36.422 25.805 1.00 10.11 A C ATOM 330 CD2 LEU A57 50.491 38.694 26.888 1.00 10.01 A C ATOM 331 C LEU A 57 52.922 34.97329.074 1.00 12.23 A C ATOM 332 O LEU A 57 53.974 34.734 28.477 1.00 9.19A O ATOM 333 N GLY A 58 52.824 34.984 30.404 1.00 11.88 A N ATOM 334 CAGLY A 58 53.969 34.719 31.249 1.00 14.46 A C ATOM 335 C GLY A 58 54.40333.278 31.089 1.00 17.50 A C ATOM 336 O GLY A 58 55.594 32.988 30.9281.00 18.69 A O ATOM 337 N GLN A 59 53.435 32.368 31.132 1.00 18.46 A NATOM 338 CA GLN A 59 53.728 30.958 30.975 1.00 19.45 A C ATOM 339 CB GLNA 59 52.463 30.128 31.161 1.00 22.28 A C ATOM 340 CG GLN A 59 51.87730.260 32.549 1.00 24.82 A C ATOM 341 CD GLN A 59 52.956 30.214 33.6191.00 25.90 A C ATOM 342 OE1 GLN A 59 53.655 29.212 33.768 1.00 26.11 A OATOM 343 NE2 GLN A 59 53.104 31.312 34.361 1.00 26.90 A N ATOM 344 C GLNA 59 54.299 30.729 29.592 1.00 19.50 A C ATOM 345 O GLN A 59 55.21829.927 29.410 1.00 20.93 A O ATOM 346 N TYR A 60 53.765 31.450 28.6161.00 17.52 A N ATOM 347 CA TYR A 60 54.239 31.315 27.252 1.00 18.33 A CATOM 348 CB TYR A 60 53.432 32.220 26.317 1.00 14.85 A C ATOM 349 CG TYRA 60 53.776 32.069 24.845 1.00 15.47 A C ATOM 350 CD1 TYR A 60 53.27931.001 24.089 1.00 14.63 A C ATOM 351 CE1 TYR A 60 53.581 30.873 22.7161.00 11.14 A C ATOM 352 CD2 TYR A 60 54.594 33.007 24.196 1.00 15.88 A CATOM 353 CE2 TYR A 60 54.902 32.886 22.826 1.00 13.26 A C ATOM 354 CZTYR A 60 54.391 31.818 22.096 1.00 12.81 A C ATOM 355 OH TYR A 60 54.67831.716 20.748 1.00 10.53 A O ATOM 356 C TYR A 60 55.728 31.664 27.1581.00 20.42 A C ATOM 357 O TYR A 60 56.557 30.793 26.871 1.00 19.39 A OATOM 358 N ILE A 61 56.069 32.928 27.418 1.00 21.45 A N ATOM 359 CA ILEA 61 57.458 33.361 27.319 1.00 23.11 A C ATOM 360 CB ILE A 61 57.62434.874 27.588 1.00 22.76 A C ATOM 361 CG2 ILE A 61 56.939 35.682 26.4991.00 23.31 A C ATOM 362 CG1 ILE A 61 57.085 35.230 28.965 1.00 22.24 A CATOM 363 CD1 ILE A 61 57.384 36.655 29.349 1.00 23.32 A C ATOM 364 C ILEA 61 58.410 32.604 28.235 1.00 25.11 A C ATOM 365 O ILE A 61 59.57332.396 27.899 1.00 25.25 A O ATOM 366 N MET A 62 57.931 32.186 29.3931.00 27.56 A N ATOM 367 CA MET A 62 58.797 31.452 30.293 1.00 30.23 A CATOM 368 CB MET A 62 58.158 31.371 31.680 1.00 32.88 A C ATOM 369 CG META 62 59.042 30.734 32.738 1.00 34.56 A C ATOM 370 SD MET A 62 58.11329.481 33.637 1.00 39.80 A S ATOM 371 CE MET A 62 58.272 28.115 32.4711.00 35.55 A C ATOM 372 C MET A 62 59.064 30.043 29.737 1.00 31.22 A CATOM 373 O MET A 62 60.218 29.656 29.513 1.00 30.54 A O ATOM 374 N THR A63 57.997 29.291 29.483 1.00 31.07 A N ATOM 375 CA THR A 63 58.15027.931 28.976 1.00 31.77 A C ATOM 376 CB THR A 63 56.775 27.264 28.6871.00 30.74 A C ATOM 377 OG1 THR A 63 56.009 28.092 27.808 1.00 31.02 A OATOM 378 CG2 THR A 63 56.004 27.044 29.982 1.00 31.19 A C ATOM 379 C THRA 63 59.023 27.823 27.724 1.00 32.00 A C ATOM 380 O THR A 63 59.89026.947 27.645 1.00 33.74 A O ATOM 381 N LYS A 64 58.797 28.697 26.7471.00 30.45 A N ATOM 382 CA LYS A 64 59.578 28.655 25.522 1.00 29.14 A CATOM 383 CB LYS A 64 58.783 29.262 24.363 1.00 28.22 A C ATOM 384 CG LYSA 64 57.581 28.437 23.947 1.00 26.54 A C ATOM 385 CD LYS A 64 56.87029.029 22.738 1.00 26.08 A C ATOM 386 CE LYS A 64 57.757 29.029 21.5141.00 26.73 A C ATOM 387 NZ LYS A 64 57.043 29.526 20.308 1.00 26.87 A NATOM 388 C LYS A 64 60.926 29.358 25.674 1.00 29.64 A C ATOM 389 O LYS A64 61.640 29.574 24.695 1.00 28.68 A O ATOM 390 N ARG A 65 61.264 29.71626.909 1.00 31.08 A N ATOM 391 CA ARG A 65 62.541 30.359 27.206 1.0031.77 A C ATOM 392 CB ARG A 65 63.624 29.278 27.281 1.00 32.61 A C ATOM393 CG ARG A 65 63.422 28.319 28.456 1.00 36.27 A C ATOM 394 CD ARG A 6564.128 26.980 28.268 1.00 38.75 A C ATOM 395 NE ARG A 65 63.951 26.10829.433 1.00 41.50 A N ATOM 396 CZ ARG A 65 64.107 24.782 29.425 1.0043.14 A C ATOM 397 NH1 ARG A 65 64.444 24.144 28.308 1.00 43.25 A N ATOM398 NH2 ARG A 65 63.923 24.087 30.540 1.00 41.34 A N ATOM 399 C ARG A 6562.931 31.442 26.193 1.00 30.62 A C ATOM 400 O ARG A 65 63.990 31.36525.562 1.00 31.11 A O ATOM 401 N LEU A 66 62.069 32.447 26.043 1.0027.85 A N ATOM 402 CA LEU A 66 62.315 33.554 25.117 1.00 24.57 A C ATOM403 CB LEU A 66 60.996 34.080 24.535 1.00 21.73 A C ATOM 404 CG LEU A 6660.195 33.192 23.588 1.00 18.58 A C ATOM 405 CD1 LEU A 66 58.917 33.89323.171 1.00 16.38 A C ATOM 406 CD2 LEU A 66 61.039 32.882 22.373 1.0016.78 A C ATOM 407 C LEU A 66 63.023 34.696 25.829 1.00 24.06 A C ATOM408 O LEU A 66 63.334 35.715 25.221 1.00 22.74 A O ATOM 409 N TYR A 6763.255 34.535 27.127 1.00 25.28 A N ATOM 410 CA TYR A 67 63.931 35.56927.900 1.00 26.02 A C ATOM 411 CB TYR A 67 63.406 35.580 29.344 1.0026.69 A C ATOM 412 CG TYR A 67 63.546 34.267 30.069 1.00 26.95 A C ATOM413 CD1 TYR A 67 64.668 34.000 30.850 1.00 27.52 A C ATOM 414 CE1 TYR A67 64.836 32.770 31.469 1.00 27.15 A C ATOM 415 CD2 TYR A 67 62.58733.269 29.928 1.00 26.57 A C ATOM 416 CE2 TYR A 67 62.742 32.031 30.5441.00 27.84 A C ATOM 417 CZ TYR A 67 63.875 31.787 31.312 1.00 27.23 A CATOM 418 OH TYR A 67 64.068 30.554 31.888 1.00 24.32 A O ATOM 419 C TYRA 67 65.443 35.347 27.857 1.00 26.40 A C ATOM 420 O TYR A 67 65.91734.216 27.828 1.00 25.07 A O ATOM 421 N ASP A 68 66.194 36.440 27.8371.00 28.69 A N ATOM 422 CA ASP A 68 67.645 36.365 27.764 1.00 31.43 A CATOM 423 CB ASP A 68 68.222 37.737 27.411 1.00 31.86 A C ATOM 424 CG ASPA 68 69.688 37.666 27.043 1.00 31.50 A C ATOM 425 OD1 ASP A 68 69.99737.105 25.972 1.00 29.47 A O ATOM 426 OD2 ASP A 68 70.526 38.154 27.8331.00 33.11 A O ATOM 427 C ASP A 68 68.313 35.848 29.035 1.00 33.44 A CATOM 428 O ASP A 68 67.871 36.128 30.152 1.00 33.12 A O ATOM 429 N GLU A69 69.399 35.105 28.845 1.00 36.37 A N ATOM 430 CA GLU A 69 70.15234.529 29.950 1.00 39.07 A C ATOM 431 CB GLU A 69 71.214 33.558 29.4201.00 42.56 A C ATOM 432 CG GLU A 69 70.627 32.277 28.851 1.00 48.37 A CATOM 433 CD GLU A 69 69.565 31.677 29.766 1.00 51.95 A C ATOM 434 OE1GLU A 69 69.848 31.514 30.974 1.00 54.24 A O ATOM 435 OE2 GLU A 6968.449 31.370 29.281 1.00 54.25 A O ATOM 436 C GLU A 69 70.810 35.56630.837 1.00 38.35 A C ATOM 437 O GLU A 69 70.695 35.506 32.057 1.0038.84 A O ATOM 438 N LYS A 70 71.504 36.519 30.235 1.00 38.42 A N ATOM439 CA LYS A 70 72.162 37.542 31.030 1.00 38.82 A C ATOM 440 CB LYS A 7073.285 38.187 30.210 1.00 41.21 A C ATOM 441 CG LYS A 70 74.320 37.14829.776 1.00 43.81 A C ATOM 442 CD LYS A 70 75.539 37.743 29.092 1.0046.99 A C ATOM 443 CE LYS A 70 76.504 36.632 28.674 1.00 47.44 A C ATOM444 NZ LYS A 70 77.794 37.156 28.143 1.00 48.71 A N ATOM 445 C LYS A 7071.121 38.554 31.494 1.00 37.27 A C ATOM 446 O LYS A 70 70.854 38.67332.690 1.00 36.92 A O ATOM 447 N GLN A 71 70.512 39.262 30.551 1.0035.72 A N ATOM 448 CA GLN A 71 69.476 40.228 30.893 1.00 33.41 A C ATOM449 CB GLN A 71 69.478 41.365 29.878 1.00 34.05 A C ATOM 450 CG GLN A 7170.781 42.116 29.818 1.00 35.15 A C ATOM 451 CD GLN A 71 70.728 43.26428.839 1.00 36.85 A C ATOM 452 OE1 GLN A 71 70.697 43.062 27.621 1.0037.63 A O ATOM 453 NE2 GLN A 71 70.701 44.482 29.364 1.00 38.20 A N ATOM454 C GLN A 71 68.115 39.512 30.898 1.00 31.00 A C ATOM 455 O GLN A 7167.412 39.466 29.883 1.00 31.06 A O ATOM 456 N GLN A 72 67.754 38.95732.051 1.00 27.04 A N ATOM 457 CA GLN A 72 66.505 38.216 32.200 1.0023.77 A C ATOM 458 CB GLN A 72 66.437 37.583 33.592 1.00 21.94 A C ATOM459 CG GLN A 72 65.172 36.783 33.877 1.00 19.04 A C ATOM 460 CD GLN A 7265.343 35.849 35.066 1.00 18.27 A C ATOM 461 OE1 GLN A 72 66.141 34.91735.018 1.00 16.43 A O ATOM 462 NE2 GLN A 72 64.600 36.098 36.137 1.0019.03 A N ATOM 463 C GLN A 72 65.231 39.003 31.936 1.00 21.79 A C ATOM464 O GLN A 72 64.255 38.438 31.464 1.00 20.39 A O ATOM 465 N HIS A 7365.234 40.300 32.226 1.00 21.17 A N ATOM 466 CA HIS A 73 64.039 41.10332.004 1.00 19.46 A C ATOM 467 CB HIS A 73 64.143 42.440 32.738 1.0019.40 A C ATOM 468 CG HIS A 73 65.242 43.328 32.244 1.00 21.41 A C ATOM469 CD2 HIS A 73 66.560 43.376 32.556 1.00 21.24 A C ATOM 470 ND1 HIS A73 65.029 44.339 31.331 1.00 22.42 A N ATOM 471 CE1 HIS A 73 66.16744.972 31.104 1.00 21.05 A C ATOM 472 NE2 HIS A 73 67.111 44.406 31.8361.00 21.25 A N ATOM 473 C HIS A 73 63.772 41.338 30.526 1.00 20.37 A CATOM 474 O HIS A 73 62.667 41.747 30.156 1.00 19.89 A O ATOM 475 N ILE A74 64.774 41.083 29.681 1.00 19.94 A N ATOM 476 CA ILE A 74 64.60741.259 28.239 1.00 19.76 A C ATOM 477 CB ILE A 74 65.942 41.605 27.5231.00 20.27 A C ATOM 478 CG2 ILE A 74 65.699 41.745 26.022 1.00 18.27 A CATOM 479 CG1 ILE A 74 66.536 42.906 28.072 1.00 22.09 A C ATOM 480 CD1ILE A 74 65.704 44.127 27.810 1.00 22.98 A C ATOM 481 C ILE A 74 64.06039.970 27.611 1.00 19.56 A C ATOM 482 O ILE A 74 64.555 38.875 27.8851.00 19.09 A O ATOM 483 N VAL A 75 63.045 40.115 26.764 1.00 17.84 A NATOM 484 CA VAL A 75 62.429 38.982 26.087 1.00 16.49 A C ATOM 485 CB VALA 75 60.918 38.928 26.388 1.00 14.74 A C ATOM 486 CG1 VAL A 75 60.24537.892 25.529 1.00 13.12 A C ATOM 487 CG2 VAL A 75 60.707 38.602 27.8521.00 15.39 A C ATOM 488 C VAL A 75 62.650 39.136 24.588 1.00 17.78 A CATOM 489 O VAL A 75 62.249 40.140 23.995 1.00 17.51 A O ATOM 490 N TYR A76 63.302 38.150 23.977 1.00 17.97 A N ATOM 491 CA TYR A 76 63.57038.201 22.544 1.00 18.96 A C ATOM 492 CB TYR A 76 64.958 37.622 22.2291.00 20.36 A C ATOM 493 CG TYR A 76 66.085 38.507 22.713 1.00 22.91 A CATOM 494 CD1 TYR A 76 66.677 38.306 23.966 1.00 22.46 A C ATOM 495 CE1TYR A 76 67.685 39.168 24.438 1.00 24.23 A C ATOM 496 CD2 TYR A 7666.525 39.587 21.942 1.00 23.56 A C ATOM 497 CE2 TYR A 76 67.532 40.45722.408 1.00 23.98 A C ATOM 498 CZ TYR A 76 68.102 40.242 23.653 1.0023.95 A C ATOM 499 OH TYR A 76 69.065 41.112 24.120 1.00 24.89 A O ATOM500 C TYR A 76 62.499 37.438 21.801 1.00 17.43 A C ATOM 501 O TYR A 7662.277 36.273 22.074 1.00 18.57 A O ATOM 502 N CYS A 77 61.841 38.09420.850 1.00 19.17 A N ATOM 503 CA CYS A 77 60.755 37.458 20.100 1.0020.17 A C ATOM 504 CB CYS A 77 59.410 37.978 20.628 1.00 18.05 A C ATOM505 SG CYS A 77 59.288 39.802 20.678 1.00 17.89 A S ATOM 506 C CYS A 7760.792 37.612 18.575 1.00 20.92 A C ATOM 507 O CYS A 77 59.817 37.29317.902 1.00 22.93 A O ATOM 508 N SER A 78 61.904 38.083 18.027 1.0021.73 A N ATOM 509 CA SER A 78 62.002 38.261 16.582 1.00 23.67 A C ATOM510 CB SER A 78 63.369 38.828 16.209 1.00 23.31 A C ATOM 511 OG SER A 7864.394 38.058 16.804 1.00 24.57 A O ATOM 512 C SER A 78 61.754 36.97815.791 1.00 24.32 A C ATOM 513 O SER A 78 61.341 37.039 14.632 1.0024.57 A O ATOM 514 N ASN A 79 62.003 35.824 16.403 1.00 23.65 A N ATOM515 CA ASN A 79 61.794 34.553 15.709 1.00 24.00 A C ATOM 516 CB ASN A 7963.081 33.709 15.694 1.00 24.75 A C ATOM 517 CG ASN A 79 64.160 34.29314.792 1.00 25.05 A C ATOM 518 OD1 ASN A 79 65.059 34.994 15.254 1.0025.04 A O ATOM 519 ND2 ASN A 79 64.064 34.015 13.495 1.00 25.70 A N ATOM520 C ASN A 79 60.668 33.742 16.343 1.00 23.53 A C ATOM 521 O ASN A 7960.753 32.512 16.451 1.00 23.89 A O ATOM 522 N ASP A 80 59.612 34.43716.747 1.00 21.43 A N ATOM 523 CA ASP A 80 58.472 33.797 17.381 1.0020.47 A C ATOM 524 CB ASP A 80 58.658 33.789 18.907 1.00 20.36 A C ATOM525 CG ASP A 80 57.567 33.022 19.626 1.00 20.02 A C ATOM 526 OD1 ASP A80 57.870 31.966 20.215 1.00 20.17 A O ATOM 527 OD2 ASP A 80 56.40433.469 19.597 1.00 20.97 A O ATOM 528 C ASP A 80 57.193 34.542 17.0241.00 20.46 A C ATOM 529 O ASP A 80 57.218 35.752 16.742 1.00 20.32 A OATOM 530 N LEU A 81 56.078 33.812 17.034 1.00 19.74 A N ATOM 531 CA LEUA 81 54.766 34.379 16.725 1.00 19.05 A C ATOM 532 CB LEU A 81 53.68533.351 17.047 1.00 20.17 A C ATOM 533 CG LEU A 81 52.228 33.802 17.0231.00 23.86 A C ATOM 534 CD1 LEU A 81 51.870 34.357 15.650 1.00 24.25 A CATOM 535 CD2 LEU A 81 51.339 32.608 17.378 1.00 24.57 A C ATOM 536 C LEUA 81 54.537 35.657 17.539 1.00 17.50 A C ATOM 537 O LEU A 81 54.02236.656 17.027 1.00 15.88 A O ATOM 538 N LEU A 82 54.948 35.604 18.8051.00 14.87 A N ATOM 539 CA LEU A 82 54.821 36.718 19.734 1.00 12.91 A CATOM 540 CB LEU A 82 55.545 36.376 21.049 1.00 11.35 A C ATOM 541 CG LEUA 82 55.603 37.457 22.138 1.00 10.75 A C ATOM 542 CD1 LEU A 82 54.18737.899 22.505 1.00 10.31 A C ATOM 543 CD2 LEU A 82 56.344 36.922 23.3541.00 9.38 A C ATOM 544 C LEU A 82 55.374 38.023 19.157 1.00 12.97 A CATOM 545 O LEU A 82 54.738 39.083 19.263 1.00 9.83 A O ATOM 546 N GLY A83 56.561 37.944 18.557 1.00 12.95 A N ATOM 547 CA GLY A 83 57.16339.129 17.980 1.00 15.06 A C ATOM 548 C GLY A 83 56.238 39.761 16.9511.00 16.77 A C ATOM 549 O GLY A 83 56.060 40.980 16.919 1.00 15.55 A OATOM 550 N ASP A 84 55.635 38.924 16.112 1.00 18.65 A N ATOM 551 CA ASPA 84 54.742 39.412 15.078 1.00 21.41 A C ATOM 552 CB ASP A 84 54.34738.272 14.141 1.00 25.41 A C ATOM 553 CG ASP A 84 55.547 37.604 13.5071.00 28.55 A C ATOM 554 OD1 ASP A 84 56.489 38.335 13.100 1.00 27.94 A OATOM 555 OD2 ASP A 84 55.539 36.354 13.412 1.00 30.30 A O ATOM 556 C ASPA 84 53.491 40.055 15.650 1.00 21.92 A C ATOM 557 O ASP A 84 53.06541.104 15.173 1.00 22.99 A O ATOM 558 N LEU A 85 52.906 39.428 16.6661.00 21.04 A N ATOM 559 CA LEU A 85 51.697 39.955 17.288 1.00 22.00 A CATOM 560 CB LEU A 85 51.092 38.915 18.240 1.00 21.98 A C ATOM 561 CG LEUA 85 50.821 37.544 17.609 1.00 23.19 A C ATOM 562 CD1 LEU A 85 50.17736.593 18.628 1.00 20.96 A C ATOM 563 CD2 LEU A 85 49.923 37.734 16.3901.00 22.60 A C ATOM 564 C LEU A 85 51.979 41.258 18.039 1.00 21.67 A CATOM 565 O LEU A 85 51.122 42.143 18.102 1.00 20.70 A O ATOM 566 N PHE A86 53.182 41.376 18.597 1.00 21.86 A N ATOM 567 CA PHE A 86 53.56542.579 19.338 1.00 20.91 A C ATOM 568 CB PHE A 86 54.531 42.224 20.4701.00 19.44 A C ATOM 569 CG PHE A 86 53.852 41.763 21.741 1.00 16.86 A CATOM 570 CD1 PHE A 86 52.508 41.381 21.742 1.00 15.83 A C ATOM 571 CD2PHE A 86 54.564 41.710 22.936 1.00 14.83 A C ATOM 572 CE1 PHE A 8651.878 40.952 22.914 1.00 14.04 A C ATOM 573 CE2 PHE A 86 53.949 41.28124.119 1.00 18.00 A C ATOM 574 CZ PHE A 86 52.596 40.902 24.105 1.0016.08 A C ATOM 575 C PHE A 86 54.211 43.604 18.416 1.00 22.21 A C ATOM576 O PHE A 86 54.233 44.797 18.714 1.00 23.98 A O ATOM 577 N GLY A 8754.728 43.140 17.284 1.00 22.25 A N ATOM 578 CA GLY A 87 55.360 44.05516.356 1.00 20.93 A C ATOM 579 C GLY A 87 56.711 44.578 16.820 1.0020.93 A C ATOM 580 O GLY A 87 57.055 45.731 16.534 1.00 22.00 A O ATOM581 N VAL A 88 57.471 43.752 17.542 1.00 18.42 A N ATOM 582 CA VAL A 8858.802 44.136 18.014 1.00 16.75 A C ATOM 583 CB VAL A 88 58.791 44.79319.420 1.00 16.55 A C ATOM 584 CG1 VAL A 88 57.951 46.054 19.405 1.0015.93 A C ATOM 585 CG2 VAL A 88 58.312 43.795 20.462 1.00 13.35 A C ATOM586 C VAL A 88 59.695 42.911 18.100 1.00 16.74 A C ATOM 587 O VAL A 8859.215 41.792 18.273 1.00 16.22 A O ATOM 588 N PRO A 89 61.013 43.11417.978 1.00 16.36 A N ATOM 589 CD PRO A 89 61.636 44.398 17.606 1.0017.10 A C ATOM 590 CA PRO A 89 62.013 42.042 18.040 1.00 16.05 A C ATOM591 CB PRO A 89 63.221 42.659 17.344 1.00 15.83 A C ATOM 592 CG PRO A 8963.124 44.096 17.747 1.00 18.07 A C ATOM 593 C PRO A 89 62.322 41.62319.476 1.00 16.21 A C ATOM 594 O PRO A 89 62.832 40.522 19.715 1.0015.73 A O ATOM 595 N SER A 90 62.009 42.505 20.425 1.00 15.13 A N ATOM596 CA SER A 90 62.243 42.232 21.843 1.00 14.02 A C ATOM 597 CB SER A 9063.738 42.128 22.130 1.00 12.56 A C ATOM 598 OG SER A 90 64.336 43.40321.986 1.00 13.74 A O ATOM 599 C SER A 90 61.652 43.350 22.702 1.0012.89 A C ATOM 600 O SER A 90 61.331 44.422 22.192 1.00 12.05 A O ATOM601 N PHE A 91 61.517 43.091 24.001 1.00 11.19 A N ATOM 602 CA PHE A 9160.972 44.069 24.926 1.00 11.45 A C ATOM 603 CB PHE A 91 59.439 44.17324.759 1.00 10.33 A C ATOM 604 CG PHE A 91 58.705 42.895 25.039 1.008.05 A C ATOM 605 CD1 PHE A 91 58.263 42.598 26.325 1.00 7.52 A C ATOM606 CD2 PHE A 91 58.496 41.961 24.024 1.00 7.09 A C ATOM 607 CE1 PHE A91 57.620 41.373 26.605 1.00 7.90 A C ATOM 608 CE2 PHE A 91 57.85940.739 24.287 1.00 7.31 A C ATOM 609 CZ PHE A 91 57.419 40.445 25.5881.00 7.15 A C ATOM 610 C PHE A 91 61.327 43.737 26.373 1.00 13.35 A CATOM 611 O PHE A 91 61.795 42.632 26.685 1.00 12.34 A O ATOM 612 N SER A92 61.102 44.708 27.254 1.00 14.78 A N ATOM 613 CA SER A 92 61.39844.547 28.667 1.00 16.74 A C ATOM 614 CB SER A 92 62.086 45.809 29.2031.00 15.00 A C ATOM 615 OG SER A 92 62.293 45.719 30.607 1.00 15.51 A OATOM 616 C SER A 92 60.148 44.254 29.499 1.00 18.09 A C ATOM 617 O SER A92 59.156 44.978 29.422 1.00 17.05 A O ATOM 618 N VAL A 93 60.206 43.18930.295 1.00 19.76 A N ATOM 619 CA VAL A 93 59.094 42.822 31.163 1.0021.70 A C ATOM 620 CB VAL A 93 59.322 41.442 31.866 1.00 22.34 A C ATOM621 CG1 VAL A 93 59.380 40.329 30.828 1.00 22.82 A C ATOM 622 CG2 VAL A93 60.606 41.462 32.683 1.00 20.47 A C ATOM 623 C VAL A 93 58.891 43.88432.239 1.00 22.48 A C ATOM 624 O VAL A 93 58.089 43.702 33.140 1.0024.56 A O ATOM 625 N LYS A 94 59.618 44.991 32.146 1.00 24.37 A N ATOM626 CA LYS A 94 59.494 46.078 33.121 1.00 26.51 A C ATOM 627 CB LYS A 9460.873 46.623 33.515 1.00 27.71 A C ATOM 628 CG LYS A 94 61.581 45.88834.659 1.00 30.96 A C ATOM 629 CD LYS A 94 62.965 46.516 34.914 1.0033.87 A C ATOM 630 CE LYS A 94 63.695 45.915 36.127 1.00 35.49 A C ATOM631 NZ LYS A 94 65.052 46.534 36.343 1.00 35.43 A N ATOM 632 C LYS A 9458.655 47.232 32.579 1.00 26.44 A C ATOM 633 O LYS A 94 58.156 48.05433.342 1.00 27.30 A O ATOM 634 N GLU A 95 58.522 47.305 31.259 1.0025.84 A N ATOM 635 CA GLU A 95 57.747 48.361 30.620 1.00 25.31 A C ATOM636 CB GLU A 95 58.317 48.651 29.231 1.00 26.45 A C ATOM 637 CG GLU A 9559.798 49.016 29.200 1.00 28.64 A C ATOM 638 CD GLU A 95 60.081 50.45229.615 1.00 30.17 A C ATOM 639 OE1 GLU A 95 59.138 51.277 29.622 1.0031.51 A O ATOM 640 OE2 GLU A 95 61.257 50.759 29.917 1.00 28.42 A O ATOM641 C GLU A 95 56.286 47.904 30.500 1.00 24.66 A C ATOM 642 O GLU A 9555.776 47.667 29.398 1.00 22.08 A O ATOM 643 N HIS A 96 55.619 47.79931.647 1.00 24.62 A N ATOM 644 CA HIS A 96 54.234 47.348 31.702 1.0026.26 A C ATOM 645 CB HIS A 96 53.694 47.452 33.132 1.00 27.90 A C ATOM646 CG HIS A 96 54.457 46.635 34.128 1.00 30.90 A C ATOM 647 CD2 HIS A96 55.784 46.395 34.257 1.00 31.52 A C ATOM 648 ND1 HIS A 96 53.84445.963 35.164 1.00 32.30 A N ATOM 649 CE1 HIS A 96 54.761 45.345 35.8871.00 32.43 A C ATOM 650 NE2 HIS A 96 55.947 45.591 35.358 1.00 32.16 A NATOM 651 C HIS A 96 53.279 48.052 30.747 1.00 26.13 A C ATOM 652 O HIS A96 52.436 47.400 30.136 1.00 26.36 A O ATOM 653 N ARG A 97 53.403 49.37030.618 1.00 25.66 A N ATOM 654 CA ARG A 97 52.522 50.123 29.728 1.0025.19 A C ATOM 655 CB ARG A 97 52.708 51.639 29.953 1.00 25.74 A C ATOM656 CG ARG A 97 51.909 52.562 29.031 1.00 24.90 A C ATOM 657 CD ARG A 9750.440 52.148 28.919 1.00 27.96 A C ATOM 658 NE ARG A 97 49.677 52.29430.157 1.00 29.51 A N ATOM 659 CZ ARG A 97 48.450 51.804 30.337 1.0029.81 A C ATOM 660 NH1 ARG A 97 47.849 51.133 29.359 1.00 31.04 A N ATOM661 NH2 ARG A 97 47.817 51.986 31.489 1.00 27.76 A N ATOM 662 C ARG A 9752.758 49.745 28.262 1.00 24.39 A C ATOM 663 O ARG A 97 51.805 49.62327.497 1.00 25.23 A O ATOM 664 N LYS A 98 54.010 49.547 27.865 1.0023.55 A N ATOM 665 CA LYS A 98 54.288 49.166 26.480 1.00 23.84 A C ATOM666 CB LYS A 98 55.800 49.167 26.195 1.00 25.86 A C ATOM 667 CG LYS A 9856.406 50.558 26.039 1.00 29.53 A C ATOM 668 CD LYS A 98 57.892 50.51225.693 1.00 31.47 A C ATOM 669 CE LYS A 98 58.519 51.909 25.794 1.0033.99 A C ATOM 670 NZ LYS A 98 59.989 51.918 25.509 1.00 34.37 A N ATOM671 C LYS A 98 53.708 47.779 26.170 1.00 23.22 A C ATOM 672 O LYS A 9853.150 47.563 25.091 1.00 21.35 A O ATOM 673 N ILE A 99 53.844 46.84627.115 1.00 21.20 A N ATOM 674 CA ILE A 99 53.323 45.496 26.938 1.0020.16 A C ATOM 675 CB ILE A 99 53.723 44.584 28.130 1.00 18.26 A C ATOM676 CG2 ILE A 99 52.911 43.290 28.108 1.00 16.58 A C ATOM 677 CG1 ILE A99 55.227 44.283 28.063 1.00 16.19 A C ATOM 678 CD1 ILE A 99 55.76443.539 29.255 1.00 12.79 A C ATOM 679 C ILE A 99 51.799 45.545 26.8021.00 21.79 A C ATOM .680 O ILE A 99 51.217 44.909 25.919 1.00 21.34 A OATOM 681 N TYR A 100 51.156 46.316 27.672 1.00 23.34 A N ATOM 682 CA TYRA 100 49.706 46.457 27.639 1.00 24.14 A C ATOM 683 CB TYR A 100 49.24347.387 28.763 1.00 25.12 A C ATOM 684 CG TYR A 100 48.602 46.651 29.9111.00 28.22 A C ATOM 685 CD1 TYR A 100 49.336 45.739 30.681 1.00 28.65 AC ATOM 686 CE1 TYR A 100 48.734 45.008 31.700 1.00 28.51 A C ATOM 687CD2 TYR A 100 47.248 46.817 30.196 1.00 28.65 A C ATOM 688 CE2 TYR A 10046.632 46.088 31.215 1.00 29.38 A C ATOM 689 CZ TYR A 100 47.380 45.18731.957 1.00 29.62 A C ATOM 690 OH TYR A 100 46.766 44.450 32.938 1.0031.04 A O ATOM 691 C TYR A 100 49.154 46.960 26.299 1.00 25.10 A C ATOM692 O TYR A 100 48.138 46.446 25.812 1.00 25.32 A O ATOM 693 N THR A 10149.798 47.956 25.693 1.00 23.58 A N ATOM 694 CA THR A 101 49.269 48.44724.434 1.00 24.54 A C ATOM 695 CB THR A 101 49.683 49.926 24.162 1.0025.48 A C ATOM 696 OG1 THR A 101 51.085 50.021 23.920 1.00 26.87 A OATOM 697 CG2 THR A 101 49.330 50.786 25.361 1.00 27.33 A C ATOM 698 CTHR A 101 49.627 47.539 23.259 1.00 23.52 A C ATOM 699 O THR A 10149.034 47.642 22.188 1.00 24.28 A O ATOM 700 N MET A 102 50.585 46.63623.450 1.00 22.80 A N ATOM 701 CA MET A 102 50.921 45.697 22.377 1.0020.40 A C ATOM 702 CB MET A 102 52.321 45.109 22.567 1.00 15.37 A C ATOM703 CG MET A 102 53.403 46.131 22.321 1.00 14.03 A C ATOM 704 SD MET A102 55.075 45.483 22.349 1.00 11.93 A S ATOM 705 CE MET A 102 55.29445.184 24.125 1.00 10.87 A C ATOM 706 C MET A 102 49.863 44.592 22.3921.00 19.65 A C ATOM 707 O MET A 102 49.528 44.017 21.356 1.00 19.47 A OATOM 708 N ILE A 103 49.338 44.319 23.580 1.00 18.90 A N ATOM 709 CA ILEA 103 48.300 43.321 23.764 1.00 21.34 A C ATOM 710 CB ILE A 103 48.13142.966 25.273 1.00 20.40 A C ATOM 711 CG2 ILE A 103 46.835 42.185 25.4971.00 19.03 A C ATOM 712 CG1 ILE A 103 49.339 42.156 25.750 1.00 19.84 AC ATOM 713 CD1 ILE A 103 49.457 42.046 27.259 1.00 18.82 A C ATOM 714 CILE A 103 46.985 43.896 23.228 1.00 24.13 A C ATOM 715 O ILE A 10346.189 43.185 22.612 1.00 24.18 A O ATOM 716 N TYR A 104 46.775 45.19223.461 1.00 27.06 A N ATOM 717 CA TYR A 104 45.566 45.886 23.016 1.0029.10 A C ATOM 718 CB TYR A 104 45.608 47.344 23.491 1.00 30.33 A C ATOM719 CG TYR A 104 45.205 47.494 24.940 1.00 30.58 A C ATOM 720 CD1 TYR A104 45.775 48.469 25.754 1.00 29.56 A C ATOM 721 CE1 TYR A 104 45.41548.584 27.095 1.00 29.30 A C ATOM 722 CD2 TYR A 104 44.259 46.638 25.5021.00 31.51 A C ATOM 723 CE2 TYR A 104 43.894 46.746 26.836 1.00 31.90 AC ATOM 724 CZ TYR A 104 44.476 47.718 27.625 1.00 30.22 A C ATOM 725 OHTYR A 104 44.113 47.801 28.947 1.00 32.03 A O ATOM 726 C TYR A 10445.321 45.824 21.510 1.00 29.49 A C ATOM 727 O TYR A 104 44.173 45.76921.067 1.00 30.07 A O ATOM 728 N ARG A 105 46.397 45.831 20.731 1.0030.44 A N ATOM 729 CA ARG A 105 46.291 45.757 19.281 1.00 31.09 A C ATOM730 CB ARG A 105 47.561 46.288 18.614 1.00 32.59 A C ATOM 731 CG ARG A105 47.624 47.803 18.491 1.00 35.48 A C ATOM 732 CD ARG A 105 48.76148.212 17.567 1.00 38.68 A C ATOM 733 NE ARG A 105 50.069 47.877 18.1291.00 41.14 A N ATOM 734 CZ ARG A 105 50.806 48.712 18.859 1.00 42.11 A CATOM 735 NH1 ARG A 105 50.368 49.942 19.117 1.00 41.27 A N ATOM 736 NH2ARG A 105 51.984 48.317 19.333 1.00 42.61 A N ATOM 737 C ARG A 10546.059 44.317 18.845 1.00 31.15 A C ATOM 738 O ARG A 105 45.919 44.04017.649 1.00 30.58 A O ATOM 739 N ASN A 106 46.029 43.403 19.814 1.0030.35 A N ATOM 740 CA ASN A 106 45.797 41.992 19.523 1.00 31.38 A C ATOM741 CB ASN A 106 46.927 41.120 20.069 1.00 30.13 A C ATOM 742 CG ASN A106 48.164 41.186 19.219 1.00 30.74 A C ATOM 743 OD1 ASN A 106 48.95742.126 19.327 1.00 30.89 A O ATOM 744 ND2 ASN A 106 48.335 40.193 18.3461.00 30.12 A N ATOM 745 C ASN A 106 44.480 41.497 20.087 1.00 32.04 A CATOM 746 O ASN A 106 44.309 40.303 20.324 1.00 30.99 A O ATOM 747 N LEUA 107 43.543 42.409 20.297 1.00 34.74 A N ATOM 748 CA LEU A 107 42.25742.009 20.834 1.00 39.45 A C ATOM 749 CB LEU A 107 42.392 41.716 22.3351.00 39.46 A C ATOM 750 CG LEU A 107 43.091 42.738 23.238 1.00 38.61 A CATOM 751 CD1 LEU A 107 42.333 44.055 23.242 1.00 38.52 A C ATOM 752 CD2LEU A 107 43.181 42.176 24.647 1.00 37.16 A C ATOM 753 C LEU A 10741.157 43.025 20.604 1.00 42.25 A C ATOM 754 O LEU A 107 41.327 43.99319.859 1.00 42.75 A O ATOM 755 N VAL A 108 40.021 42.778 21.245 1.0046.00 A N ATOM 756 CA VAL A 108 38.859 43.653 21.163 1.00 49.43 A C ATOM757 CB VAL A 108 37.823 43.129 20.149 1.00 48.08 A C ATOM 758 CG1 VAL A108 37.030 44.290 19.594 1.00 48.02 A C ATOM 759 CG2 VAL A 108 38.50942.343 19.040 1.00 46.78 A C ATOM 760 C VAL A 108 38.216 43.671 22.5551.00 53.19 A C ATOM 761 O VAL A 108 37.589 42.690 22.966 1.00 52.78 A OATOM 762 N VAL A 109 38.386 44.778 23.280 1.00 57.34 A N ATOM 763 CA VALA 109 37.829 44.921 24.630 1.00 61.10 A C ATOM 764 CB VAL A 109 38.03546.351 25.191 1.00 61.46 A C ATOM 765 CG VAL A 109 37.715 46.367 26.6831.00 61.63 A C ATOM 766 CG2 VAL A 109 39.458 46.830 24.928 1.00 62.28 AC ATOM 767 C VAL A 109 36.326 44.631 24.662 1.00 63.47 A C ATOM 768 OVAL A 109 35.527 45.410 24.131 1.00 63.73 A O ATOM 769 N VAL A 11035.947 43.518 25.293 1.00 65.46 A N ATOM 770 CA VAL A 110 34.542 43.12725.392 1.00 67.21 A C ATOM 771 CB VAL A 110 34.366 41.919 26.365 1.0066.97 A C ATOM 772 CG1 VAL A 110 32.939 41.374 26.283 1.00 67.04 A CATOM 773 CG2 VAL A 110 35.371 40.825 26.033 1.00 66.24 A C ATOM 774 CVAL A 110 33.697 44.309 25.898 1.00 69.20 A C ATOM 775 O VAL A 11034.287 45.249 26.488 1.00 69.94 A O ATOM 776 OXT VAL A 110 32.456 44.28425.705 1.00 70.58 A O ATOM 777 C1 CID A 1 55.200 42.184 33.980 1.0021.13 INH1 C ATOM 778 C2 CID A 1 54.610 43.160 33.125 1.00 21.31 INH1 CATOM 779 C3 CID A 1 53.194 43.197 32.974 1.00 21.72 INH1 C ATOM 780 C4CID A 1 52.372 42.284 33.667 1.00 21.74 INH1 C ATOM 781 C5 CID A 152.953 41.311 34.515 1.00 21.42 INH1 C ATOM 782 C6 CID A 1 54.387 41.23834.692 1.00 22.49 INH1 C ATOM 783 C7 CID A 1 55.092 40.168 35.590 1.0023.66 INH1 C ATOM 784 C8 CID A 1 54.267 39.594 36.800 1.00 25.69 INH1 CATOM 785 O1 CID A 1 54.677 38.706 37.544 1.00 29.01 INH1 O ATOM 786 O2CID A 1 53.229 40.347 37.224 1.00 30.64 INH1 O ATOM 787 N1 CID A 155.591 39.038 34.674 1.00 19.99 INH1 N ATOM 788 C9 CID A 1 54.593 38.35233.801 1.00 18.95 INH1 C ATOM 789 C10 CID A 1 54.631 38.684 32.289 1.0018.04 INH1 C ATOM 790 C11 CID A 1 55.647 39.488 31.665 1.00 17.01 INH1 CATOM 791 C12 CID A 1 55.614 39.775 30.286 1.00 16.97 INH1 C ATOM 792 C13CID A 1 54.563 39.261 29.507 1.00 17.64 INH1 C ATOM 793 CL1 CID A 154.498 39.606 27.865 1.00 14.96 INH1CL ATOM 794 C14 CID A 1 53.55038.464 30.083 1.00 18.96 INH1 C ATOM 795 C15 CID A 1 53.586 38.18031.458 1.00 17.55 INH1 C ATOM 796 C16 CID A 1 54.559 36.817 34.087 1.0018.08 INH1 C ATOM 797 O3 CID A 1 53.499 36.278 34.423 1.00 18.82 INH1 OATOM 798 N2 CID A 1 55.695 36.036 33.977 1.00 16.78 INH1 N ATOM 799 C17CID A 1 57.002 36.408 33.618 1.00 17.03 INH1 C ATOM 800 C18 CID A 157.616 37.644 33.943 1.00 16.70 INH1 C ATOM 801 C19 CID A 1 56.97238.734 34.754 1.00 19.45 INH1 C ATOM 802 O4 CID A 1 57.728 39.367 35.5321.00 18.52 INH1 O ATOM 803 C20 CID A 1 58.948 37.897 33.495 1.00 17.26INH1 C ATOM 804 C21 CID A 1 59.660 36.940 32.750 1.00 18.91 INH1 C ATOM805 I1 CID A 1 61.599 37.431 32.161 1.00 19.64 INH1 I ATOM 806 C22 CID A1 59.069 35.711 32.436 1.00 17.86 INH1 C ATOM 807 C23 CID A 1 57.74235.435 32.859 1.00 17.23 INH1 C ATOM 808 CL2 CID A 1 52.462 44.35431.946 1.00 20.99 INH1CL ATOM 809 CL3 CID A 1 59.915 34.517 31.548 1.0020.31 INH1CL END

[0227] TABLE 3 Superimposed: trigonal and tetragonal crystal formsREMARK Superimposed on /xray1/hmdm2/PDB/M338437.pdb REMARK The 19 atomshave an RMS distance of 0.249 A REMARK RMS delta B = 6.724 A2 REMARKEstimated RMSD for 2 random proteins = 5.398 A REMARK Relative RMSD =0.04621 REMARK Normalised RMSD (100) = 1.471 A REMARK coordinates fromrestrained individual B-factor refinement REMARK refinement resolution:25 − 2.6 A REMARK starting r = 0.2563 free_r = 0.2787 REMARK final r =0.2553 free_r = 0.2761 REMARK B rmsd for bonded mainchain atoms = 1.483target = 1.5 REMARK B rmsd for bonded sidechain atoms = 1.740 target =2.0 REMARK B rmsd for angle mainchain atoms = 2.593 target = 2.0 REMARKB rmsd for angle sidechain atoms = 2.780 target = 2.5 REMARK rweight =0.1000 (with wa = 3.71696) REMARK target = mlf steps = 30 REMARK sg =P4(3)2(1)2 a = 54.3 b = 54.3 c = 83.3 alpha = 90 beta = 90 gamma = 90REMARK parameter file 1: MSI_CNX_TOPPAR: protein_rep.param REMARKparameter file 2: ../cid.par REMARK molecular structure file:recycle.psf REMARK input coordinates: anneal_9.pdb REMARK reflectionfile = ../M876273_2_P43212.cv REMARK ncs = none REMARK B-correctionresolution: 6.0 − 2.6 REMARK initial B-factor correction applied tofobs: REMARK B11 = −1.189 B22 = −1.189 B33 = 2.379 REMARK B12 = 0.000B13 = 0.000 B23 = 0.000 REMARK B-factor correction applied to coordinatearray B: −0.119 REMARK bulk solvent: (Mask) density level = 0.341945e/A{circumflex over ( )}3, B-factor = 22.3925 A{circumflex over ( )}2REMARK reflections with |Fobs|/sigma_F < 0.0 rejected REMARK reflectionswith |Fobs| > 10000 * rms(Fobs) rejected REMARK theoretical total numberof refl.in resol.range: 4173 (100.0%) REMARK number of unobservedreflections (no entry or |F| = 0): 9 (0.2%) REMARK number of reflectionsrejected: 0 (0.0%) REMARK total number of reflections used: 4164 (99.8%)REMARK number of reflections in working set: 3737 (89.6%) REMARK numberof reflections in test set: 427 (10.2%) REMARK FILENAME =“bindividual.pdb” REMARK Written by CNX VERSION: 2000.12 ATOM 1 C GLY A16 48.607 19.990 25.187 1.00 68.15 A ATOM 2 O GLY A 16 48.239 21.10624.797 1.00 68.22 A ATOM 3 N GLY A 16 47.838 17.646 24.774 1.00 67.11 AATOM 4 CA GLY A 16 47.594 18.911 25.537 1.00 67.90 A ATOM 5 N SER A 1749.889 19.652 25.332 1.00 67.05 A ATOM 6 CA SER A 17 50.986 20.56825.025 1.00 64.73 A ATOM 7 CB SER A 17 51.581 21.155 26.312 1.00 65.01 AATOM 8 OG SER A 17 50.639 21.978 26.989 1.00 63.84 A ATOM 9 C SER A 1752.053 19.794 24.258 1.00 62.82 A ATOM 10 O SER A 17 52.921 20.38223.611 1.00 62.75 A ATOM 11 N GLN A 18 51.970 18.468 24.343 1.00 60.52 AATOM 12 CA GLN A 18 52.895 17.577 23.647 1.00 57.89 A ATOM 13 CB GLN A18 52.794 16.161 24.210 1.00 57.50 A ATOM 14 CG GLN A 18 53.480 15.95525.534 1.00 57.38 A ATOM 15 CD GLN A 18 53.377 14.514 25.999 1.00 58.16A ATOM 16 OE1 GLN A 18 53.614 13.581 25.228 1.00 56.84 A ATOM 17 NE2 GLNA 18 53.027 14.327 27.268 1.00 58.44 A ATOM 18 C GLN A 18 52.532 17.52922.169 1.00 55.87 A ATOM 19 O GLN A 18 53.378 17.267 21.312 1.00 55.83 AATOM 20 N ILE A 19 51.256 17.781 21.889 1.00 52.87 A ATOM 21 CA ILE A 1950.727 17.763 20.532 1.00 50.05 A ATOM 22 CB ILE A 19 49.408 16.94020.476 1.00 48.06 A ATOM 23 CG2 ILE A 19 48.886 16.873 19.053 1.00 48.17A ATOM 24 CG1 ILE A 19 49.638 15.526 21.020 1.00 45.44 A ATOM 25 CD1 ILEA 19 50.552 14.677 20.180 1.00 43.26 A ATOM 26 C ILE A 19 50.443 19.19420.066 1.00 49.72 A ATOM 27 O ILE A 19 50.014 20.036 20.856 1.00 49.03 AATOM 28 N PRO A 20 50.702 19.490 18.777 1.00 49.57 A ATOM 29 CD PRO A 2051.486 18.667 17.841 1.00 49.58 A ATOM 30 CA PRO A 20 50.469 20.82218.209 1.00 49.39 A ATOM 31 CB PRO A 20 51.058 20.705 16.808 1.00 48.67A ATOM 32 CG PRO A 20 52.153 19.717 16.991 1.00 48.94 A ATOM 33 C PRO A20 48.982 21.187 18.171 1.00 49.97 A ATOM 34 O PRO A 20 48.138 20.35817.819 1.00 50.18 A ATOM 35 N ALA A 21 48.672 22.429 18.534 1.00 49.45 AATOM 36 CA ALA A 21 47.296 22.913 18.540 1.00 49.81 A ATOM 37 CB ALA A21 47.270 24.405 18.880 1.00 49.85 A ATOM 38 C ALA A 21 46.613 22.67017.189 1.00 49.65 A ATOM 39 O ALA A 21 45.483 22.179 17.128 1.00 49.33 AATOM 40 N SER A 22 47.302 23.022 16.107 1.00 48.76 A ATOM 41 CA SER A 2246.753 22.830 14.774 1.00 47.75 A ATOM 42 CB SER A 22 47.823 23.13413.721 1.00 47.15 A ATOM 43 OG SER A 22 49.001 22.382 13.964 1.00 48.43A ATOM 44 C SER A 22 46.254 21.391 14.628 1.00 46.34 A ATOM 45 O SER A22 45.195 21.143 14.045 1.00 45.89 A ATOM 46 N GLU A 23 47.012 20.44515.172 1.00 44.68 A ATOM 47 CA GLU A 23 46.632 19.041 15.098 1.00 43.25A ATOM 48 CB GLU A 23 47.804 18.149 15.472 1.00 42.16 A ATOM 49 CG GLU A23 47.513 16.684 15.303 1.00 40.86 A ATOM 50 CD GLU A 23 48.777 15.86615.250 1.00 40.82 A ATOM 51 OE1 GLU A 23 49.695 16.146 16.045 1.00 41.52A ATOM 52 OE2 GLU A 23 48.856 14.942 14.418 1.00 41.56 A ATOM 53 C GLU A23 45.453 18.760 16.013 1.00 42.01 A ATOM 54 O GLU A 23 44.505 18.08715.625 1.00 42.32 A ATOM 55 N GLN A 24 45.512 19.278 17.229 1.00 41.29 AATOM 56 CA GLN A 24 44.413 19.089 18.154 1.00 42.07 A ATOM 57 CB GLN A24 44.666 19.872 19.450 1.00 41.13 A ATOM 58 CG GLN A 24 45.643 19.18020.391 1.00 42.57 A ATOM 59 CD GLN A 24 45.950 19.981 21.650 1.00 43.91A ATOM 60 OE1 GLN A 24 45.068 20.622 22.233 1.00 44.84 A ATOM 61 NE2 GLNA 24 47.205 19.931 22.085 1.00 43.17 A ATOM 62 C GLN A 24 43.140 19.58917.475 1.00 43.32 A ATOM 63 O GLN A 24 42.035 19.139 17.790 1.00 43.40 AATOM 64 N GLU A 25 43.310 20.505 16.521 1.00 44.62 A ATOM 65 CA GLU A 2542.183 21.095 15.795 1.00 44.92 A ATOM 66 CB GLU A 25 42.507 22.54315.406 1.00 48.33 A ATOM 67 CG GLU A 25 43.121 23.398 16.516 1.00 53.04A ATOM 68 CD GLU A 25 42.283 23.449 17.787 1.00 55.79 A ATOM 69 OE1 GLUA 25 42.680 24.180 18.720 1.00 57.11 A ATOM 70 OE2 GLU A 25 41.23622.766 17.864 1.00 57.84 A ATOM 71 C GLU A 25 41.731 20.336 14.541 1.0042.75 A ATOM 72 O GLU A 25 40.616 20.547 14.059 1.00 42.62 A ATOM 73 NTHR A 26 42.587 19.467 14.008 1.00 40.18 A ATOM 74 CA THR A 26 42.23718.692 12.814 1.00 38.13 A ATOM 75 CB THR A 26 43.254 17.547 12.563 1.0037.92 A ATOM 76 OG1 THR A 26 44.589 18.059 12.647 1.00 37.02 A ATOM 77CG2 THR A 26 43.047 16.942 11.187 1.00 36.76 A ATOM 78 C THR A 26 40.84718.074 12.983 1.00 36.79 A ATOM 79 O THR A 26 40.511 17.574 14.054 1.0036.47 A ATOM 80 N LEU A 27 40.036 18.128 11.931 1.00 36.63 A ATOM 81 CALEU A 27 38.686 17.559 11.973 1.00 35.77 A ATOM 82 CB LEU A 27 37.73918.336 11.052 1.00 36.16 A ATOM 83 CG LEU A 27 36.264 18.393 11.488 1.0038.05 A ATOM 84 CD1 LEU A 27 36.120 19.331 12.692 1.00 36.12 A ATOM 85CD2 LEU A 27 35.394 18.895 10.328 1.00 38.41 A ATOM 86 C LEU A 27 38.79616.110 11.505 1.00 34.52 A ATOM 87 O LEU A 27 39.467 15.818 10.513 1.0033.72 A ATOM 88 N VAL A 28 38.135 15.204 12.218 1.00 33.53 A ATOM 89 CAVAL A 28 38.214 13.787 11.886 1.00 32.88 A ATOM 90 CB VAL A 28 39.20713.071 12.850 1.00 32.86 A ATOM 91 CG1 VAL A 28 40.592 13.685 12.7241.00 31.21 A ATOM 92 CG2 VAL A 28 38.726 13.204 14.292 1.00 33.40 A ATOM93 C VAL A 28 36.876 13.044 11.919 1.00 31.29 A ATOM 94 O VAL A 2835.910 13.501 12.527 1.00 30.99 A ATOM 95 N ARG A 29 36.841 11.89711.248 1.00 29.80 A ATOM 96 CA ARG A 29 35.655 11.054 11.198 1.00 28.94A ATOM 97 CB ARG A 29 35.174 10.876 9.762 1.00 33.14 A ATOM 98 CG ARG A29 34.296 11.991 9.254 1.00 38.54 A ATOM 99 CD ARG A 29 34.036 11.8307.767 1.00 43.39 A ATOM 100 NE ARG A 29 33.084 12.828 7.296 1.00 47.28 AATOM 101 CZ ARG A 29 31.772 12.741 7.477 1.00 49.30 A ATOM 102 NH1 ARG A29 31.257 11.691 8.110 1.00 49.75 A ATOM 103 NH2 ARG A 29 30.978 13.7167.049 1.00 50.29 A ATOM 104 C ARG A 29 35.994 9.688 11.762 1.00 26.43 AATOM 105 O ARG A 29 36.680 8.895 11.110 1.00 25.41 A ATOM 106 N PRO A 3035.528 9.397 12.989 1.00 24.69 A ATOM 107 CD PRO A 30 34.749 10.28513.869 1.00 23.43 A ATOM 108 CA PRO A 30 35.784 8.108 13.647 1.00 23.00A ATOM 109 CB PRO A 30 35.223 8.308 15.053 1.00 21.72 A ATOM 110 CG PROA 30 35.147 9.792 15.218 1.00 22.48 A ATOM 111 C PRO A 30 35.023 7.00112.923 1.00 22.20 A ATOM 112 O PRO A 30 33.945 7.243 12.382 1.00 22.27 AATOM 113 N LYS A 31 35.580 5.796 12.910 1.00 21.65 A ATOM 114 CA LYS A31 34.909 4.658 12.286 1.00 19.51 A ATOM 115 CB LYS A 31 35.901 3.51912.056 1.00 19.68 A ATOM 116 CG LYS A 31 37.058 3.899 11.150 1.00 20.82A ATOM 117 CD LYS A 31 38.006 2.724 10.976 1.00 22.74 A ATOM 118 CE LYSA 31 39.161 3.040 10.019 1.00 20.68 A ATOM 119 NZ LYS A 31 40.000 1.8189.826 1.00 21.75 A ATOM 120 C LYS A 31 33.795 4.225 13.249 1.00 18.36 AATOM 121 O LYS A 31 33.793 4.605 14.422 1.00 17.38 A ATOM 122 N PRO A 3232.848 3.406 12.774 1.00 17.15 A ATOM 123 CD PRO A 32 32.886 2.66511.501 1.00 16.30 A ATOM 124 CA PRO A 32 31.729 2.939 13.595 1.00 16.70A ATOM 125 CB PRO A 32 31.178 1.775 12.778 1.00 16.53 A ATOM 126 CG PROA 32 31.463 2.200 11.378 1.00 16.35 A ATOM 127 C PRO A 32 32.023 2.54815.048 1.00 16.73 A ATOM 128 O PRO A 32 31.343 3.014 15.967 1.00 17.12 AATOM 129 N LEU A 33 33.016 1.692 15.264 1.00 14.62 A ATOM 130 CA LEU A33 33.324 1.265 16.619 1.00 13.59 A ATOM 131 CB LEU A 33 34.332 0.11016.594 1.00 15.36 A ATOM 132 CG LEU A 33 33.787 −1.301 16.896 1.00 13.91A ATOM 133 CD1 LEU A 33 32.250 −1.320 16.992 1.00 13.51 A ATOM 134 CD2LEU A 33 34.270 −2.239 15.822 1.00 8.87 A ATOM 135 C LEU A 33 33.8002.397 17.516 1.00 13.14 A ATOM 136 O LEU A 33 33.281 2.560 18.608 1.0012.37 A ATOM 137 N LEU A 34 34.786 3.176 17.082 1.00 14.58 A ATOM 138 CALEU A 34 35.238 4.305 17.895 1.00 13.84 A ATOM 139 CB LEU A 34 36.4305.015 17.246 1.00 13.73 A ATOM 140 CG LEU A 34 36.892 6.346 17.861 1.0010.65 A ATOM 141 CD1 LEU A 34 37.437 6.120 19.247 1.00 9.38 A ATOM 142CD2 LEU A 34 37.951 6.971 16.986 1.00 11.47 A ATOM 143 C LEU A 34 34.0735.294 18.042 1.00 15.75 A ATOM 144 O LEU A 34 33.877 5.874 19.103 1.0016.50 A ATOM 145 N LEU A 35 33.294 5.484 16.979 1.00 17.07 A ATOM 146 CALEU A 35 32.152 6.395 17.046 1.00 18.94 A ATOM 147 CB LEU A 35 31.4406.482 15.690 1.00 16.70 A ATOM 148 CG LEU A 35 30.311 7.514 15.602 1.0014.51 A ATOM 149 CD1 LEU A 35 30.880 8.904 15.849 1.00 13.73 A ATOM 150CD2 LEU A 35 29.646 7.453 14.237 1.00 13.01 A ATOM 151 C LEU A 35 31.1515.968 18.136 1.00 20.66 A ATOM 152 O LEU A 35 30.494 6.823 18.742 1.0021.36 A ATOM 153 N LYS A 36 31.039 4.661 18.385 1.00 20.77 A ATOM 154 CALYS A 36 30.135 4.152 19.415 1.00 22.28 A ATOM 155 CB LYS A 36 29.9382.637 19.298 1.00 23.62 A ATOM 156 CG LYS A 36 28.839 2.214 18.330 1.0028.23 A ATOM 157 CD LYS A 36 28.007 1.043 18.889 1.00 30.56 A ATOM 158CE LYS A 36 28.853 −0.202 19.171 1.00 32.43 A ATOM 159 NZ LYS A 3628.037 −1.385 19.580 1.00 31.16 A ATOM 160 C LYS A 36 30.668 4.47120.807 1.00 23.86 A ATOM 161 O LYS A 36 29.901 4.855 21.700 1.00 26.24 AATOM 162 N LEU A 37 31.971 4.301 21.006 1.00 23.16 A ATOM 163 CA LEU A37 32.558 4.608 22.308 1.00 24.32 A ATOM 164 CB LEU A 37 34.085 4.51522.263 1.00 25.18 A ATOM 165 CG LEU A 37 34.708 3.137 22.100 1.00 27.89A ATOM 166 CD1 LEU A 37 36.217 3.232 22.302 1.00 28.24 A ATOM 167 CD2LEU A 37 34.095 2.198 23.119 1.00 28.07 A ATOM 168 C LEU A 37 32.1796.027 22.725 1.00 23.05 A ATOM 169 O LEU A 37 31.758 6.268 23.854 1.0020.34 A ATOM 170 N LEU A 38 32.338 6.956 21.788 1.00 23.36 A ATOM 171 CALEU A 38 32.055 8.359 22.022 1.00 24.58 A ATOM 172 CB LEU A 38 32.4939.198 20.819 1.00 21.20 A ATOM 173 CG LEU A 38 33.886 8.937 20.238 1.0019.16 A ATOM 174 CD1 LEU A 38 34.126 9.914 19.106 1.00 16.77 A ATOM 175CD2 LEU A 38 34.966 9.092 21.306 1.00 18.04 A ATOM 176 C LEU A 38 30.5818.602 22.302 1.00 27.20 A ATOM 177 O LEU A 38 30.236 9.411 23.162 1.0029.33 A ATOM 178 N LYS A 39 29.702 7.908 21.590 1.00 28.13 A ATOM 179 CALYS A 39 28.283 8.119 21.815 1.00 29.30 A ATOM 180 CB LYS A 39 27.4677.488 20.686 1.00 28.63 A ATOM 181 CG LYS A 39 27.777 8.127 19.347 1.0028.78 A ATOM 182 CD LYS A 39 26.776 7.761 18.276 1.00 27.98 A ATOM 183CE LYS A 39 27.154 8.415 16.960 1.00 26.89 A ATOM 184 NZ LYS A 39 26.0748.260 15.959 1.00 27.11 A ATOM 185 C LYS A 39 27.840 7.587 23.169 1.0030.33 A ATOM 186 O LYS A 39 26.931 8.141 23.789 1.00 31.74 A ATOM 187 NSER A 40 28.495 6.531 23.644 1.00 30.27 A ATOM 188 CA SER A 40 28.1485.948 24.939 1.00 29.62 A ATOM 189 CB SER A 40 28.995 4.691 25.213 1.0028.81 A ATOM 190 OG SER A 40 30.349 5.002 25.520 1.00 26.68 A ATOM 191 CSER A 40 28.340 6.960 26.073 1.00 29.48 A ATOM 192 O SER A 40 27.7456.822 27.139 1.00 29.59 A ATOM 193 N VAL A 41 29.170 7.974 25.843 1.0030.35 A ATOM 194 CA VAL A 41 29.432 9.002 26.854 1.00 30.76 A ATOM 195CB VAL A 41 30.944 9.047 27.267 1.00 30.64 A ATOM 196 CG1 VAL A 4131.298 7.826 28.115 1.00 29.00 A ATOM 197 CG2 VAL A 41 31.834 9.10826.030 1.00 28.32 A ATOM 198 C VAL A 41 29.013 10.407 26.413 1.00 32.02A ATOM 199 O VAL A 41 29.742 11.380 26.628 1.00 31.05 A ATOM 200 N GLY A42 27.845 10.509 25.779 1.00 33.77 A ATOM 201 CA GLY A 42 27.355 11.81125.360 1.00 36.14 A ATOM 202 C GLY A 42 27.488 12.202 23.901 1.00 38.10A ATOM 203 O GLY A 42 26.516 12.669 23.310 1.00 40.02 A ATOM 204 N ALA A43 28.675 12.035 23.318 1.00 38.97 A ATOM 205 CA ALA A 43 28.897 12.40321.919 1.00 39.16 A ATOM 206 CB ALA A 43 30.124 11.678 21.370 1.00 38.86A ATOM 207 C ALA A 43 27.678 12.101 21.054 1.00 39.94 A ATOM 208 O ALA A43 26.968 11.120 21.284 1.00 40.04 A ATOM 209 N GLN A 44 27.435 12.95220.061 1.00 41.54 A ATOM 210 CA GLN A 44 26.294 12.769 19.173 1.00 42.78A ATOM 211 CB GLN A 44 25.018 13.232 19.881 1.00 45.11 A ATOM 212 CG GLNA 44 25.132 14.598 20.546 1.00 47.61 A ATOM 213 CD GLN A 44 24.02614.838 21.568 1.00 49.56 A ATOM 214 OE1 GLN A 44 23.986 15.884 22.2271.00 49.35 A ATOM 215 NE2 GLN A 44 23.124 13.866 21.708 1.00 49.08 AATOM 216 C GLN A 44 26.434 13.470 17.821 1.00 42.23 A ATOM 217 O GLN A44 25.520 14.171 17.375 1.00 42.70 A ATOM 218 N LYS A 45 27.585 13.27117.180 1.00 40.15 A ATOM 219 CA LYS A 45 27.871 13.845 15.870 1.00 37.90A ATOM 220 CB LYS A 45 28.802 15.050 15.980 1.00 39.62 A ATOM 221 CG LYSA 45 28.359 16.141 16.933 1.00 41.18 A ATOM 222 CD LYS A 45 29.19717.399 16.713 1.00 42.96 A ATOM 223 CE LYS A 45 30.690 17.093 16.7321.00 45.64 A ATOM 224 NZ LYS A 45 31.535 18.294 16.451 1.00 48.08 A ATOM225 C LYS A 45 28.590 12.774 15.071 1.00 36.61 A ATOM 226 O LYS A 4528.771 11.657 15.553 1.00 36.82 A ATOM 227 N ASP A 46 29.012 13.12113.859 1.00 34.69 A ATOM 228 CA ASP A 46 29.736 12.187 13.006 1.00 33.64A ATOM 229 CB ASP A 46 29.089 12.074 11.617 1.00 35.21 A ATOM 230 CG ASPA 46 27.726 11.403 11.652 1.00 36.60 A ATOM 231 OD1 ASP A 46 27.55710.427 12.417 1.00 35.63 A ATOM 232 OD2 ASP A 46 26.830 11.846 10.8991.00 36.79 A ATOM 233 C ASP A 46 31.174 12.651 12.830 1.00 32.32 A ATOM234 O ASP A 46 32.071 11.843 12.578 1.00 33.06 A ATOM 235 N THR A 4731.388 13.959 12.950 1.00 30.30 A ATOM 236 CA THR A 47 32.718 14.54312.790 1.00 27.52 A ATOM 237 CB THR A 47 32.762 15.556 11.620 1.00 28.37A ATOM 238 OG1 THR A 47 31.586 16.375 11.650 1.00 28.12 A ATOM 239 CG2THR A 47 32.846 14.836 10.287 1.00 28.11 A ATOM 240 C THR A 47 33.13815.254 14.059 1.00 25.47 A ATOM 241 O THR A 47 32.307 15.811 14.781 1.0024.25 A ATOM 242 N TYR A 48 34.441 15.227 14.324 1.00 24.41 A ATOM 243CA TYR A 48 34.997 15.844 15.517 1.00 24.02 A ATOM 244 CB TYR A 4835.073 14.830 16.681 1.00 23.01 A ATOM 245 CG TYR A 48 33.769 14.16217.077 1.00 22.29 A ATOM 246 CD1 TYR A 48 33.259 13.087 16.347 1.0021.16 A ATOM 247 CE1 TYR A 48 32.030 12.499 16.685 1.00 20.20 A ATOM 248CD2 TYR A 48 33.021 14.632 18.166 1.00 21.34 A ATOM 249 CE2 TYR A 4831.801 14.050 18.510 1.00 20.29 A ATOM 250 CZ TYR A 48 31.312 12.99017.762 1.00 19.58 A ATOM 251 OH TYR A 48 30.092 12.442 18.066 1.00 20.11A ATOM 252 C TYR A 48 36.412 16.340 15.265 1.00 24.38 A ATOM 253 O TYR A48 37.080 15.913 14.318 1.00 22.46 A ATOM 254 N THR A 49 36.855 17.24516.130 1.00 24.50 A ATOM 255 CA THR A 49 38.221 17.744 16.092 1.00 25.25A ATOM 256 CB THR A 49 38.333 19.134 16.762 1.00 26.57 A ATOM 257 OG1THR A 49 37.583 19.131 17.989 1.00 26.24 A ATOM 258 CG2 THR A 49 37.79620.235 15.829 1.00 24.91 A ATOM 259 C THR A 49 38.932 16.699 16.965 1.0025.63 A ATOM 260 O THR A 49 38.307 16.088 17.833 1.00 24.71 A ATOM 261 NMET A 50 40.216 16.466 16.740 1.00 26.39 A ATOM 262 CA MET A 50 40.90715.470 17.545 1.00 26.62 A ATOM 263 CB MET A 50 42.373 15.388 17.1501.00 25.74 A ATOM 264 CG MET A 50 42.589 14.617 15.865 1.00 25.74 A ATOM265 SD MET A 50 42.250 12.852 16.072 1.00 24.59 A ATOM 266 CE MET A 5043.816 12.259 16.701 1.00 22.52 A ATOM 267 C MET A 50 40.785 15.76019.031 1.00 28.23 A ATOM 268 O MET A 50 40.843 14.848 19.847 1.00 29.67A ATOM 269 N LYS A 51 40.594 17.026 19.388 1.00 28.66 A ATOM 270 CA LYSA 51 40.467 17.377 20.794 1.00 29.11 A ATOM 271 CB LYS A 51 40.57318.891 20.980 1.00 32.66 A ATOM 272 CG LYS A 51 40.687 19.300 22.4451.00 37.67 A ATOM 273 CD LYS A 51 40.874 20.801 22.628 1.00 41.07 A ATOM274 CE LYS A 51 40.948 21.164 24.116 1.00 42.73 A ATOM 275 NZ LYS A 5141.069 22.639 24.365 1.00 45.66 A ATOM 276 C LYS A 51 39.159 16.86221.406 1.00 28.29 A ATOM 277 O LYS A 51 39.143 16.411 22.557 1.00 28.36A ATOM 278 N GLU A 52 38.064 16.929 20.647 1.00 25.71 A ATOM 279 CA GLUA 52 36.775 16.451 21.144 1.00 23.60 A ATOM 280 CB GLU A 52 35.63816.866 20.199 1.00 24.83 A ATOM 281 CG GLU A 52 35.407 18.369 20.0841.00 27.15 A ATOM 282 CD GLU A 52 34.383 18.732 18.998 1.00 29.01 A ATOM283 OE1 GLU A 52 34.598 18.371 17.818 1.00 29.80 A ATOM 284 OE2 GLU A 5233.364 19.380 19.320 1.00 29.19 A ATOM 285 C GLU A 52 36.785 14.92121.293 1.00 21.67 A ATOM 286 O GLU A 52 36.069 14.370 22.132 1.00 20.99A ATOM 287 N VAL A 53 37.578 14.235 20.473 1.00 18.55 A ATOM 288 CA VALA 53 37.659 12.782 20.559 1.00 16.46 A ATOM 289 CB VAL A 53 38.46112.163 19.371 1.00 17.02 A ATOM 290 CG1 VAL A 53 38.749 10.682 19.6451.00 15.92 A ATOM 291 CG2 VAL A 53 37.659 12.282 18.075 1.00 14.61 AATOM 292 C VAL A 53 38.350 12.471 21.879 1.00 15.15 A ATOM 293 O VAL A53 37.788 11.784 22.735 1.00 13.10 A ATOM 294 N LEU A 54 39.570 12.98822.037 1.00 15.42 A ATOM 295 CA LEU A 54 40.328 12.814 23.273 1.00 14.74A ATOM 296 CB LEU A 54 41.548 13.736 23.309 1.00 12.86 A ATOM 297 CG LEUA 54 42.871 13.232 22.730 1.00 13.00 A ATOM 298 CD1 LEU A 54 43.13211.821 23.250 1.00 14.60 A ATOM 299 CD2 LEU A 54 42.833 13.235 21.2321.00 11.73 A ATOM 300 C LEU A 54 39.439 13.153 24.467 1.00 15.13 A ATOM301 O LEU A 54 39.515 12.511 25.514 1.00 16.88 A ATOM 302 N PHE A 5538.588 14.156 24.309 1.00 14.69 A ATOM 303 CA PHE A 55 37.710 14.55625.397 1.00 16.41 A ATOM 304 CB PHE A 55 36.904 15.803 25.028 1.00 18.34A ATOM 305 CG PHE A 55 35.899 16.171 26.069 1.00 20.72 A ATOM 306 CD1PHE A 55 36.278 16.916 27.184 1.00 20.24 A ATOM 307 CD2 PHE A 55 34.59615.672 26.001 1.00 20.52 A ATOM 308 CE1 PHE A 55 35.376 17.153 28.2231.00 20.60 A ATOM 309 CE2 PHE A 55 33.687 15.901 27.034 1.00 21.73 AATOM 310 CZ PHE A 55 34.078 16.644 28.149 1.00 20.59 A ATOM 311 C PHE A55 36.739 13.473 25.838 1.00 16.04 A ATOM 312 O PHE A 55 36.661 13.14827.025 1.00 17.31 A ATOM 313 N TYR A 56 35.978 12.941 24.886 1.00 16.71A ATOM 314 CA TYR A 56 34.996 11.892 25.165 1.00 17.61 A ATOM 315 CB TYRA 56 34.136 11.624 23.930 1.00 18.45 A ATOM 316 CG TYR A 56 33.14212.723 23.671 1.00 21.48 A ATOM 317 CD1 TYR A 56 32.090 12.951 24.5651.00 23.38 A ATOM 318 CE1 TYR A 56 31.182 13.987 24.363 1.00 25.16 AATOM 319 CD2 TYR A 56 33.266 13.562 22.557 1.00 22.53 A ATOM 320 CE2 TYRA 56 32.363 14.610 22.339 1.00 23.90 A ATOM 321 CZ TYR A 56 31.32214.816 23.251 1.00 26.15 A ATOM 322 OH TYR A 56 30.425 15.846 23.0681.00 26.91 A ATOM 323 C TYR A 56 35.709 10.628 25.567 1.00 16.78 A ATOM324 O TYR A 56 35.189 9.810 26.329 1.00 18.53 A ATOM 325 N LEU A 5736.910 10.472 25.028 1.00 15.78 A ATOM 326 CA LEU A 57 37.743 9.32525.327 1.00 13.13 A ATOM 327 CB LEU A 57 38.982 9.382 24.437 1.00 11.95A ATOM 328 CG LEU A 57 39.277 8.207 23.497 1.00 13.21 A ATOM 329 CD1 LEUA 57 38.000 7.628 22.913 1.00 10.11 A ATOM 330 CD2 LEU A 57 40.224 8.69022.402 1.00 10.01 A ATOM 331 C LEU A 57 38.108 9.418 26.821 1.00 12.23 AATOM 332 O LEU A 57 38.197 8.403 27.515 1.00 9.19 A ATOM 333 N GLY A 5838.291 10.649 27.300 1.00 11.88 A ATOM 334 CA GLY A 58 38.620 10.87728.691 1.00 14.46 A ATOM 335 C GLY A 58 37.448 10.490 29.567 1.00 17.50A ATOM 336 O GLY A 58 37.613 9.800 30.579 1.00 18.69 A ATOM 337 N GLN A59 36.257 10.934 29.178 1.00 18.46 A ATOM 338 CA GLN A 59 35.059 10.61429.929 1.00 19.45 A ATOM 339 CB GLN A 59 33.850 11.322 29.328 1.00 22.28A ATOM 340 CG GLN A 59 33.967 12.829 29.389 1.00 24.82 A ATOM 341 CD GLNA 59 34.506 13.295 30.732 1.00 25.90 A ATOM 342 OE1 GLN A 59 33.88313.079 31.771 1.00 26.11 A ATOM 343 NE2 GLN A 59 35.679 13.928 30.7141.00 26.90 A ATOM 344 C GLN A 59 34.850 9.115 29.898 1.00 19.50 A ATOM345 O GLN A 59 34.441 8.511 30.893 1.00 20.93 A ATOM 346 N TYR A 6035.148 8.510 28.757 1.00 17.52 A ATOM 347 CA TYR A 60 34.991 7.07528.617 1.00 18.33 A ATOM 348 CB TYR A 60 35.359 6.636 27.198 1.00 14.85A ATOM 349 CG TYR A 60 35.120 5.162 26.914 1.00 15.47 A ATOM 350 CD1 TYRA 60 33.838 4.676 26.632 1.00 14.63 A ATOM 351 CE1 TYR A 60 33.620 3.31126.347 1.00 11.14 A ATOM 352 CD2 TYR A 60 36.184 4.247 26.913 1.00 15.88A ATOM 353 CE2 TYR A 60 35.974 2.882 26.631 1.00 13.26 A ATOM 354 CZ TYRA 60 34.691 2.425 26.349 1.00 12.81 A ATOM 355 OH TYR A 60 34.494 1.08926.052 1.00 10.53 A ATOM 356 C TYR A 60 35.862 6.332 29.635 1.00 20.42 AATOM 357 O TYR A 60 35.339 5.672 30.541 1.00 19.39 A ATOM 358 N ILE A 6137.184 6.453 29.501 1.00 21.45 A ATOM 359 CA ILE A 61 38.092 5.75330.401 1.00 23.11 A ATOM 360 CB ILE A 61 39.576 5.969 30.026 1.00 22.76A ATOM 361 CG2 ILE A 61 39.879 5.334 28.680 1.00 23.31 A ATOM 362 CG1ILE A 61 39.913 7.453 30.028 1.00 22.24 A ATOM 363 CD1 ILE A 61 41.3857.709 29.846 1.00 23.32 A ATOM 364 C ILE A 61 37.911 6.115 31.869 1.0025.11 A ATOM 365 O ILE A 61 38.112 5.284 32.751 1.00 25.25 A ATOM 366 NMET A 62 37.531 7.350 32.145 1.00 27.56 A ATOM 367 CA MET A 62 37.3357.737 33.527 1.00 30.23 A ATOM 368 CB MET A 62 37.237 9.260 33.632 1.0032.88 A ATOM 369 CG MET A 62 37.161 9.783 35.056 1.00 34.56 A ATOM 370SD MET A 62 35.808 10.961 35.203 1.00 39.80 A ATOM 371 CE MET A 6234.440 9.803 35.399 1.00 35.55 A ATOM 372 C MET A 62 36.066 7.073 34.0871.00 31.22 A ATOM 373 O MET A 62 36.118 6.340 35.083 1.00 30.54 A ATOM374 N THR A 63 34.934 7.302 33.428 1.00 31.07 A ATOM 375 CA THR A 6333.673 6.735 33.896 1.00 31.77 A ATOM 376 CB THR A 63 32.495 7.07432.939 1.00 30.74 A ATOM 377 OG1 THR A 63 32.818 6.664 31.608 1.00 31.02A ATOM 378 CG2 THR A 63 32.206 8.569 32.956 1.00 31.19 A ATOM 379 C THRA 63 33.710 5.221 34.115 1.00 32.00 A ATOM 380 O THR A 63 33.230 4.73035.142 1.00 33.74 A ATOM 381 N LYS A 64 34.265 4.481 33.159 1.00 30.45 AATOM 382 CA LYS A 64 34.331 3.035 33.288 1.00 29.14 A ATOM 383 CB LYS A64 34.397 2.380 31.905 1.00 28.22 A ATOM 384 CG LYS A 64 33.121 2.52331.099 1.00 26.54 A ATOM 385 CD LYS A 64 33.198 1.785 29.769 1.00 26.08A ATOM 386 CE LYS A 64 33.343 0.293 29.967 1.00 26.73 A ATOM 387 NZ LYSA 64 33.332 −0.444 28.675 1.00 26.87 A ATOM 388 C LYS A 64 35.509 2.58634.151 1.00 29.64 A ATOM 389 O LYS A 64 35.824 1.398 34.223 1.00 28.68 AATOM 390 N ARG A 65 36.160 3.548 34.799 1.00 31.08 A ATOM 391 CA ARG A65 37.279 3.259 35.691 1.00 31.77 A ATOM 392 CB ARG A 65 36.720 2.80037.041 1.00 32.61 A ATOM 393 CG ARG A 65 35.955 3.907 37.771 1.00 36.27A ATOM 394 CD ARG A 65 34.975 3.376 38.812 1.00 38.75 A ATOM 395 NE ARGA 65 34.298 4.465 39.521 1.00 41.50 A ATOM 396 CZ ARG A 65 33.149 4.34340.190 1.00 43.14 A ATOM 397 NH1 ARG A 65 32.519 3.173 40.251 1.00 43.25A ATOM 398 NH2 ARG A 65 32.622 5.396 40.799 1.00 41.34 A ATOM 399 C ARGA 65 38.254 2.216 35.131 1.00 30.62 A ATOM 400 O ARG A 65 38.488 1.17335.750 1.00 31.11 A ATOM 401 N LEU A 66 38.817 2.505 33.958 1.00 27.85 AATOM 402 CA LEU A 66 39.773 1.606 33.309 1.00 24.57 A ATOM 403 CB LEU A66 39.657 1.699 31.781 1.00 21.73 A ATOM 404 CG LEU A 66 38.392 1.18531.099 1.00 18.58 A ATOM 405 CD1 LEU A 66 38.477 1.413 29.603 1.00 16.38A ATOM 406 CD2 LEU A 66 38.240 −0.289 31.393 1.00 16.78 A ATOM 407 C LEUA 66 41.195 1.960 33.712 1.00 24.06 A ATOM 408 O LEU A 66 42.146 1.31233.286 1.00 22.74 A ATOM 409 N TYR A 67 41.343 3.008 34.515 1.00 25.28 AATOM 410 CA TYR A 67 42.665 3.428 34.962 1.00 26.02 A ATOM 411 CB TYR A67 42.703 4.953 35.146 1.00 26.69 A ATOM 412 CG TYR A 67 41.675 5.49236.106 1.00 26.95 A ATOM 413 CD1 TYR A 67 41.983 5.673 37.452 1.00 27.52A ATOM 414 CE1 TYR A 67 41.025 6.108 38.356 1.00 27.15 A ATOM 415 CD2TYR A 67 40.378 5.764 35.683 1.00 26.57 A ATOM 416 CE2 TYR A 67 39.4076.202 36.579 1.00 27.84 A ATOM 417 CZ TYR A 67 39.738 6.368 37.919 1.0027.23 A ATOM 418 OH TYR A 67 38.780 6.753 38.826 1.00 24.32 A ATOM 419 CTYR A 67 43.032 2.700 36.256 1.00 26.40 A ATOM 420 O TYR A 67 42.1782.422 37.091 1.00 25.07 A ATOM 421 N ASP A 68 44.311 2.381 36.403 1.0028.69 A ATOM 422 CA ASP A 68 44.785 1.659 37.574 1.00 31.43 A ATOM 423CB ASP A 68 46.199 1.130 37.326 1.00 31.86 A ATOM 424 CG ASP A 68 46.6350.139 38.382 1.00 31.50 A ATOM 425 OD1 ASP A 68 46.072 −0.975 38.4091.00 29.47 A ATOM 426 OD2 ASP A 68 47.524 0.482 39.192 1.00 33.11 A ATOM427 C ASP A 68 44.770 2.474 38.864 1.00 33.44 A ATOM 428 O ASP A 6845.033 3.679 38.866 1.00 33.12 A ATOM 429 N GLU A 69 44.477 1.789 39.9661.00 36.37 A ATOM 430 CA GLU A 69 44.415 2.416 41.279 1.00 39.07 A ATOM431 CB GLU A 69 43.852 1.432 42.311 1.00 42.56 A ATOM 432 CG GLU A 6942.371 1.145 42.123 1.00 48.37 A ATOM 433 CD GLU A 69 41.564 2.42041.901 1.00 51.95 A ATOM 434 OE1 GLU A 69 41.715 3.364 42.708 1.00 54.24A ATOM 435 OE2 GLU A 69 40.782 2.480 40.921 1.00 54.25 A ATOM 436 C GLUA 69 45.750 2.946 41.759 1.00 38.35 A ATOM 437 O GLU A 69 45.845 4.08442.207 1.00 38.84 A ATOM 438 N LYS A 70 46.788 2.128 41.672 1.00 38.42 AATOM 439 CA LYS A 70 48.096 2.575 42.118 1.00 38.82 A ATOM 440 CB LYS A70 48.982 1.360 42.412 1.00 41.21 A ATOM 441 CG LYS A 70 48.362 0.47143.491 1.00 43.81 A ATOM 442 CD LYS A 70 49.261 −0.669 43.940 1.00 46.99A ATOM 443 CE LYS A 70 48.551 −1.515 44.999 1.00 47.44 A ATOM 444 NZ LYSA 70 49.436 −2.552 45.599 1.00 48.71 A ATOM 445 C LYS A 70 48.694 3.49341.058 1.00 37.27 A ATOM 446 O LYS A 70 48.890 4.684 41.300 1.00 36.92 AATOM 447 N GLN A 71 48.958 2.950 39.876 1.00 35.72 A ATOM 448 CA GLN A71 49.496 3.755 38.787 1.00 33.41 A ATOM 449 CB GLN A 71 50.372 2.88837.891 1.00 34.05 A ATOM 450 CG GLN A 71 51.544 2.272 38.608 1.00 35.15A ATOM 451 CD GLN A 71 52.414 1.462 37.678 1.00 36.85 A ATOM 452 OE1 GLNA 71 52.026 0.382 37.220 1.00 37.63 A ATOM 453 NE2 GLN A 71 53.596 1.98337.377 1.00 38.20 A ATOM 454 C GLN A 71 48.326 4.349 37.986 1.00 31.00 AATOM 455 O GLN A 71 47.855 3.759 37.007 1.00 31.06 A ATOM 456 N GLN A 7247.865 5.521 38.411 1.00 27.04 A ATOM 457 CA GLN A 72 46.737 6.19237.771 1.00 23.77 A ATOM 458 CB GLN A 72 46.355 7.443 38.567 1.00 21.94A ATOM 459 CG GLN A 72 45.189 8.240 37.996 1.00 19.04 A ATOM 460 CD GLNA 72 44.592 9.192 39.022 1.00 18.27 A ATOM 461 OE1 GLN A 72 44.040 8.75840.029 1.00 16.43 A ATOM 462 NE2 GLN A 72 44.704 10.491 38.773 1.0019.03 A ATOM 463 C GLN A 72 46.926 6.558 36.307 1.00 21.79 A ATOM 464 OGLN A 72 45.965 6.558 35.552 1.00 20.39 A ATOM 465 N HIS A 73 48.1556.859 35.899 1.00 21.17 A ATOM 466 CA HIS A 73 48.395 7.227 34.510 1.0019.46 A ATOM 467 CB HIS A 73 49.768 7.880 34.355 1.00 19.40 A ATOM 468CG HIS A 73 50.918 6.974 34.667 1.00 21.41 A ATOM 469 CD2 HIS A 7351.514 6.660 35.843 1.00 21.24 A ATOM 470 ND1 HIS A 73 51.613 6.29033.692 1.00 22.42 A ATOM 471 CE1 HIS A 73 52.588 5.596 34.254 1.00 21.05A ATOM 472 NE2 HIS A 73 52.548 5.804 35.559 1.00 21.25 A ATOM 473 C HISA 73 48.273 6.037 33.572 1.00 20.37 A ATOM 474 O HIS A 73 48.166 6.21932.355 1.00 19.89 A ATOM 475 N ILE A 74 48.290 4.823 34.128 1.00 19.94 AATOM 476 CA ILE A 74 48.158 3.618 33.310 1.00 19.76 A ATOM 477 CB ILE A74 48.869 2.389 33.941 1.00 20.27 A ATOM 478 CG2 ILE A 74 48.667 1.16533.052 1.00 18.27 A ATOM 479 CG1 ILE A 74 50.368 2.655 34.118 1.00 22.09A ATOM 480 CD1 ILE A 74 51.121 2.838 32.833 1.00 22.98 A ATOM 481 C ILEA 74 46.676 3.261 33.133 1.00 19.56 A ATOM 482 O ILE A 74 45.911 3.24534.099 1.00 19.09 A ATOM 483 N VAL A 75 46.287 2.968 31.895 1.00 17.84 AATOM 484 CA VAL A 75 44.913 2.603 31.578 1.00 16.49 A ATOM 485 CB VAL A75 44.335 3.551 30.508 1.00 14.74 A ATOM 486 CG1 VAL A 75 42.999 3.05330.027 1.00 13.12 A ATOM 487 CG2 VAL A 75 44.189 4.941 31.092 1.00 15.39A ATOM 488 C VAL A 75 44.901 1.172 31.058 1.00 17.78 A ATOM 489 O VAL A75 45.568 0.858 30.070 1.00 17.51 A ATOM 490 N TYR A 76 44.155 0.30031.731 1.00 17.97 A ATOM 491 CA TYR A 76 44.077 −1.098 31.319 1.00 18.96A ATOM 492 CB TYR A 76 44.029 −2.024 32.544 1.00 20.36 A ATOM 493 CG TYRA 76 45.341 −2.070 33.294 1.00 22.91 A ATOM 494 CD1 TYR A 76 45.582−1.227 34.385 1.00 22.46 A ATOM 495 CE1 TYR A 76 46.827 −1.231 35.0441.00 24.23 A ATOM 496 CD2 TYR A 76 46.371 −2.920 32.878 1.00 23.56 AATOM 497 CE2 TYR A 76 47.621 −2.929 33.530 1.00 23.98 A ATOM 498 CZ TYRA 76 47.840 −2.083 34.606 1.00 23.95 A ATOM 499 OH TYR A 76 49.073−2.071 35.223 1.00 24.89 A ATOM 500 C TYR A 76 42.856 −1.303 30.454 1.0017.43 A ATOM 501 O TYR A 76 41.753 −0.999 30.870 1.00 18.57 A ATOM 502 NCYS A 77 43.052 −1.833 29.250 1.00 19.17 A ATOM 503 CA CYS A 77 41.940−2.034 28.318 1.00 20.17 A ATOM 504 CB CYS A 77 41.984 −0.939 27.2421.00 18.05 A ATOM 505 SG CYS A 77 43.606 −0.777 26.414 1.00 17.89 A ATOM506 C CYS A 77 41.853 −3.405 27.639 1.00 20.92 A ATOM 507 O CYS A 7741.084 −3.577 26.698 1.00 22.93 A ATOM 508 N SER A 78 42.619 −4.37928.110 1.00 21.73 A ATOM 509 CA SER A 78 42.590 −5.706 27.504 1.00 23.67A ATOM 510 CB SER A 78 43.569 −6.636 28.216 1.00 23.31 A ATOM 511 OG SERA 78 43.353 −6.594 29.612 1.00 24.57 A ATOM 512 C SER A 78 41.202 −6.34427.497 1.00 24.32 A ATOM 513 O SER A 78 40.917 −7.189 26.647 1.00 24.57A ATOM 514 N ASN A 79 40.343 −5.949 28.432 1.00 23.65 A ATOM 515 CA ASNA 79 38.996 −6.517 28.492 1.00 24.00 A ATOM 516 CB ASN A 79 38.716−7.139 29.872 1.00 24.75 A ATOM 517 CG ASN A 79 39.516 −8.411 30.1201.00 25.05 A ATOM 518 OD1 ASN A 79 40.571 −8.380 30.753 1.00 25.04 AATOM 519 ND2 ASN A 79 39.021 −9.534 29.608 1.00 25.70 A ATOM 520 C ASN A79 37.928 −5.472 28.186 1.00 23.53 A ATOM 521 O ASN A 79 36.857 −5.45528.806 1.00 23.89 A ATOM 522 N ASP A 80 38.222 −4.611 27.219 1.00 21.43A ATOM 523 CA ASP A 80 37.304 −3.554 26.831 1.00 20.47 A ATOM 524 CB ASPA 80 37.609 −2.277 27.631 1.00 20.36 A ATOM 525 CG ASP A 80 36.608−1.170 27.370 1.00 20.02 A ATOM 526 OD1 ASP A 80 35.855 −0.817 28.2991.00 20.17 A ATOM 527 OD2 ASP A 80 36.567 −0.656 26.236 1.00 20.97 AATOM 528 C ASP A 80 37.439 −3.270 25.341 1.00 20.46 A ATOM 529 O ASP A80 38.505 −3.492 24.744 1.00 20.32 A ATOM 530 N LEU A 81 36.350 −2.78324.746 1.00 19.74 A ATOM 531 CA LEU A 81 36.317 −2.448 23.323 1.00 19.05A ATOM 532 CB LEU A 81 35.019 −1.709 23.010 1.00 20.17 A ATOM 533 CG LEUA 81 34.870 −1.058 21.638 1.00 23.86 A ATOM 534 CD1 LEU A 81 35.022−2.102 20.539 1.00 24.25 A ATOM 535 CD2 LEU A 81 33.500 −0.381 21.5631.00 24.57 A ATOM 536 C LEU A 81 37.522 −1.576 22.957 1.00 17.50 A ATOM537 O LEU A 81 38.155 −1.766 21.913 1.00 15.88 A ATOM 538 N LEU A 8237.831 −0.633 23.846 1.00 14.87 A ATOM 539 CA LEU A 82 38.944 0.29023.679 1.00 12.91 A ATOM 540 CB LEU A 82 39.117 1.125 24.961 1.00 11.35A ATOM 541 CG LEU A 82 40.296 2.107 25.024 1.00 10.75 A ATOM 542 CD1 LEUA 82 40.216 3.088 23.855 1.00 10.31 A ATOM 543 CD2 LEU A 82 40.284 2.83926.357 1.00 9.38 A ATOM 544 C LEU A 82 40.252 −0.430 23.344 1.00 12.97 AATOM 545 O LEU A 82 40.992 −0.012 22.439 1.00 9.83 A ATOM 546 N GLY A 8340.538 −1.503 24.080 1.00 12.95 A ATOM 547 CA GLY A 83 41.756 −2.24823.834 1.00 15.06 A ATOM 548 C GLY A 83 41.816 −2.728 22.391 1.00 16.77A ATOM 549 O GLY A 83 42.853 −2.635 21.732 1.00 15.55 A ATOM 550 N ASP A84 40.691 −3.233 21.894 1.00 18.65 A ATOM 551 CA ASP A 84 40.631 −3.73620.534 1.00 21.41 A ATOM 552 CB ASP A 84 39.294 −4.432 20.288 1.00 25.41A ATOM 553 CG ASP A 84 39.043 −5.561 21.263 1.00 28.55 A ATOM 554 OD1ASP A 84 40.004 −6.324 21.550 1.00 27.94 A ATOM 555 OD2 ASP A 84 37.887−5.685 21.730 1.00 30.30 A ATOM 556 C ASP A 84 40.830 −2.640 19.503 1.0021.92 A ATOM 557 O ASP A 84 41.547 −2.838 18.525 1.00 22.99 A ATOM 558 NLEU A 85 40.197 −1.491 19.719 1.00 21.04 A ATOM 559 CA LEU A 85 40.314−0.373 18.790 1.00 22.00 A ATOM 560 CB LEU A 85 39.286 0.713 19.133 1.0021.98 A ATOM 561 CG LEU A 85 37.835 0.226 19.211 1.00 23.19 A ATOM 562CD1 LEU A 85 36.884 1.393 19.514 1.00 20.96 A ATOM 563 CD2 LEU A 8537.472 −0.450 17.892 1.00 22.60 A ATOM 564 C LEU A 85 41.727 0.21418.799 1.00 21.67 A ATOM 565 O LEU A 85 42.216 0.686 17.770 1.00 20.70 AATOM 566 N PHE A 86 42.381 0.173 19.958 1.00 21.86 A ATOM 567 CA PHE A86 43.740 0.702 20.086 1.00 20.91 A ATOM 568 CB PHE A 86 43.965 1.26421.491 1.00 19.44 A ATOM 569 CG PHE A 86 43.487 2.688 21.671 1.00 16.86A ATOM 570 CD1 PHE A 86 42.626 3.282 20.744 1.00 15.83 A ATOM 571 CD2PHE A 86 43.899 3.431 22.773 1.00 14.83 A ATOM 572 CE1 PHE A 86 42.1814.597 20.908 1.00 14.04 A ATOM 573 CE2 PHE A 86 43.461 4.749 22.955 1.0018.00 A ATOM 574 CZ PHE A 86 42.597 5.334 22.013 1.00 16.08 A ATOM 575 CPHE A 86 44.774 −0.377 19.794 1.00 22.21 A ATOM 576 O PHE A 86 45.917−0.080 19.447 1.00 23.98 A ATOM 577 N GLY A 87 44.370 −1.635 19.923 1.0022.25 A ATOM 578 CA GLY A 87 45.298 −2.717 19.664 1.00 20.93 A ATOM 579C GLY A 87 46.363 −2.894 20.735 1.00 20.93 A ATOM 580 O GLY A 87 47.499−3.266 20.417 1.00 22.00 A ATOM 581 N VAL A 88 46.015 −2.621 21.994 1.0018.42 A ATOM 582 CA VAL A 88 46.947 −2.787 23.110 1.00 16.75 A ATOM 583CB VAL A 88 47.763 −1.506 23.426 1.00 16.55 A ATOM 584 CG1 VAL A 8848.589 −1.098 22.223 1.00 15.93 A ATOM 585 CG2 VAL A 88 46.836 −0.39423.892 1.00 13.35 A ATOM 586 C VAL A 88 46.185 −3.155 24.371 1.00 16.74A ATOM 587 O VAL A 88 45.010 −2.822 24.518 1.00 16.22 A ATOM 588 N PRO A89 46.854 −3.851 25.300 1.00 16.36 A ATOM 589 CD PRO A 89 48.202 −4.42025.116 1.00 17.10 A ATOM 590 CA PRO A 89 46.269 −4.283 26.574 1.00 16.05A ATOM 591 CB PRO A 89 47.181 −5.427 27.000 1.00 15.83 A ATOM 592 CG PROA 89 48.517 −4.975 26.501 1.00 18.07 A ATOM 593 C PRO A 89 46.232 −3.15527.604 1.00 16.21 A ATOM 594 O PRO A 89 45.462 −3.210 28.570 1.00 15.73A ATOM 595 N SER A 90 47.066 −2.138 27.391 1.00 15.13 A ATOM 596 CA SERA 90 47.131 −0.988 28.292 1.00 14.02 A ATOM 597 CB SER A 90 47.652−1.409 29.663 1.00 12.56 A ATOM 598 OG SER A 90 49.018 −1.764 29.5601.00 13.74 A ATOM 599 C SER A 90 48.060 0.083 27.719 1.00 12.89 A ATOM600 O SER A 90 48.833 −0.191 26.803 1.00 12.05 A ATOM 601 N PHE A 9147.978 1.293 28.267 1.00 11.19 A ATOM 602 CA PHE A 91 48.807 2.39727.816 1.00 11.45 A ATOM 603 CB PHE A 91 48.290 2.943 26.466 1.00 10.33A ATOM 604 CG PHE A 91 46.891 3.480 26.518 1.00 8.05 A ATOM 605 CD1 PHEA 91 46.655 4.817 26.828 1.00 7.52 A ATOM 606 CD2 PHE A 91 45.800 2.63826.304 1.00 7.09 A ATOM 607 CE1 PHE A 91 45.338 5.315 26.932 1.00 7.90 AATOM 608 CE2 PHE A 91 44.485 3.118 26.404 1.00 7.31 A ATOM 609 CZ PHE A91 44.256 4.467 26.721 1.00 7.15 A ATOM 610 C PHE A 91 48.869 3.51728.852 1.00 13.35 A ATOM 611 O PHE A 91 48.090 3.547 29.817 1.00 12.34 AATOM 612 N SER A 92 49.807 4.437 28.644 1.00 14.78 A ATOM 613 CA SER A92 49.996 5.558 29.548 1.00 16.74 A ATOM 614 CB SER A 92 51.493 5.76929.811 1.00 15.00 A ATOM 615 OG SER A 92 51.712 6.925 30.610 1.00 15.51A ATOM 616 C SER A 92 49.384 6.854 29.012 1.00 18.09 A ATOM 617 O SER A92 49.653 7.257 27.881 1.00 17.05 A ATOM 618 N VAL A 93 48.562 7.50229.835 1.00 19.76 A ATOM 619 CA VAL A 93 47.941 8.765 29.456 1.00 21.70A ATOM 620 CB VAL A 93 46.883 9.242 30.506 1.00 22.34 A ATOM 621 CG1 VALA 93 45.727 8.253 30.571 1.00 22.82 A ATOM 622 CG2 VAL A 93 47.520 9.39331.880 1.00 20.47 A ATOM 623 C VAL A 93 49.001 9.852 29.311 1.00 22.48 AATOM 624 O VAL A 93 48.672 11.011 29.120 1.00 24.56 A ATOM 625 N LYS A94 50.272 9.479 29.402 1.00 24.37 A ATOM 626 CA LYS A 94 51.369 10.44129.268 1.00 26.51 A ATOM 627 CB LYS A 94 52.454 10.190 30.323 1.00 27.71A ATOM 628 CG LYS A 94 52.235 10.866 31.682 1.00 30.96 A ATOM 629 CD LYSA 94 53.375 10.491 32.648 1.00 33.87 A ATOM 630 CE LYS A 94 53.29811.223 33.998 1.00 35.49 A ATOM 631 NZ LYS A 94 54.414 10.825 34.9301.00 35.43 A ATOM 632 C LYS A 94 52.014 10.375 27.886 1.00 26.44 A ATOM633 O LYS A 94 52.693 11.309 27.469 1.00 27.30 A ATOM 634 N GLU A 9551.821 9.260 27.190 1.00 25.84 A ATOM 635 CA GLU A 95 52.386 9.07625.859 1.00 25.31 A ATOM 636 CB GLU A 95 52.648 7.590 25.612 1.00 26.45A ATOM 637 CG GLU A 95 53.540 6.907 26.643 1.00 28.64 A ATOM 638 CD GLUA 95 55.022 7.198 26.457 1.00 30.17 A ATOM 639 OE1 GLU A 95 55.415 7.65725.359 1.00 31.51 A ATOM 640 OE2 GLU A 95 55.798 6.948 27.407 1.00 28.42A ATOM 641 C GLU A 95 51.394 9.612 24.818 1.00 24.66 A ATOM 642 O GLU A95 50.809 8.851 24.037 1.00 22.08 A ATOM 643 N HIS A 96 51.225 10.93224.808 1.00 24.62 A ATOM 644 CA HIS A 96 50.295 11.590 23.900 1.00 26.26A ATOM 645 CB HIS A 96 50.410 13.113 24.027 1.00 27.90 A ATOM 646 CG HISA 96 50.114 13.629 25.400 1.00 30.90 A ATOM 647 CD2 HIS A 96 50.42213.138 26.625 1.00 31.52 A ATOM 648 ND1 HIS A 96 49.432 14.807 25.6211.00 32.30 A ATOM 649 CE1 HIS A 96 49.333 15.018 26.922 1.00 32.43 AATOM 650 NE2 HIS A 96 49.926 14.019 27.554 1.00 32.16 A ATOM 651 C HIS A96 50.420 11.193 22.435 1.00 26.13 A ATOM 652 O HIS A 96 49.408 11.00621.764 1.00 26.36 A ATOM 653 N ARG A 97 51.648 11.067 21.938 1.00 25.66A ATOM 654 CA ARG A 97 51.857 10.696 20.540 1.00 25.19 A ATOM 655 CB ARGA 97 53.344 10.864 20.161 1.00 25.74 A ATOM 656 CG ARG A 97 53.73410.433 18.746 1.00 24.90 A ATOM 657 CD ARG A 97 52.779 10.981 17.6841.00 27.96 A ATOM 658 NE ARG A 97 52.817 12.435 17.532 1.00 29.51 A ATOM659 CZ ARG A 97 51.931 13.132 16.820 1.00 29.81 A ATOM 660 NH1 ARG A 9750.936 12.508 16.197 1.00 31.04 A ATOM 661 NH2 ARG A 97 52.037 14.45116.722 1.00 27.76 A ATOM 662 C ARG A 97 51.371 9.269 20.265 1.00 24.39 AATOM 663 O ARG A 97 50.775 9.012 19.222 1.00 25.23 A ATOM 664 N LYS A 9851.605 8.344 21.189 1.00 23.55 A ATOM 665 CA LYS A 98 51.145 6.97020.983 1.00 23.84 A ATOM 666 CB LYS A 98 51.678 6.034 22.082 1.00 25.86A ATOM 667 CG LYS A 98 53.151 5.669 21.933 1.00 29.53 A ATOM 668 CD LYSA 98 53.621 4.689 23.004 1.00 31.47 A ATOM 669 CE LYS A 98 55.149 4.54322.978 1.00 33.99 A ATOM 670 NZ LYS A 98 55.672 3.627 24.040 1.00 34.37A ATOM 671 C LYS A 98 49.611 6.908 20.955 1.00 23.22 A ATOM 672 O LYS A98 49.031 6.190 20.136 1.00 21.35 A ATOM 673 N ILE A 99 48.963 7.65821.849 1.00 21.20 A ATOM 674 CA ILE A 99 47.507 7.689 21.909 1.00 20.16A ATOM 675 CB ILE A 99 47.017 8.541 23.112 1.00 18.26 A ATOM 676 CG2 ILEA 99 45.526 8.843 22.980 1.00 16.58 A ATOM 677 CG1 ILE A 99 47.306 7.79324.421 1.00 16.19 A ATOM 678 CD1 ILE A 99 47.022 8.589 25.665 1.00 12.79A ATOM 679 C ILE A 99 46.949 8.256 20.601 1.00 21.79 A ATOM 680 O ILE A99 46.008 7.710 20.019 1.00 21.34 A ATOM 681 N TYR A 100 47.543 9.34820.133 1.00 23.34 A ATOM 682 CA TYR A 100 47.113 9.978 18.891 1.00 24.14A ATOM 683 CB TYR A 100 47.961 11.220 18.611 1.00 25.12 A ATOM 684 CGTYR A 100 47.228 12.508 18.883 1.00 28.22 A ATOM 685 CD1 TYR A 10046.799 12.833 20.177 1.00 28.65 A ATOM 686 CE1 TYR A 100 46.065 13.98920.423 1.00 28.51 A ATOM 687 CD2 TYR A 100 46.908 13.379 17.843 1.0028.65 A ATOM 688 CE2 TYR A 100 46.170 14.541 18.077 1.00 29.38 A ATOM689 CZ TYR A 100 45.752 14.835 19.366 1.00 29.62 A ATOM 690 OH TYR A 10045.002 15.961 19.589 1.00 31.04 A ATOM 691 C TYR A 100 47.149 9.04817.671 1.00 25.10 A ATOM 692 O TYR A 100 46.216 9.055 16.857 1.00 25.32A ATOM 693 N THR A 101 48.206 8.251 17.524 1.00 23.58 A ATOM 694 CA THRA 101 48.252 7.383 16.362 1.00 24.54 A ATOM 695 CB THR A 101 49.7147.004 15.972 1.00 25.48 A ATOM 696 OG1 THR A 101 50.297 6.160 16.9621.00 26.87 A ATOM 697 CG2 THR A 101 50.552 8.262 15.841 1.00 27.33 AATOM 698 C THR A 101 47.376 6.143 16.527 1.00 23.52 A ATOM 699 O THR A101 47.074 5.459 15.553 1.00 24.28 A ATOM 700 N MET A 102 46.949 5.85117.753 1.00 22.80 A ATOM 701 CA MET A 102 46.052 4.711 17.956 1.00 20.40A ATOM 702 CB MET A 102 46.081 4.229 19.408 1.00 15.37 A ATOM 703 CG META 102 47.385 3.556 19.756 1.00 14.03 A ATOM 704 SD MET A 102 47.4372.805 21.384 1.00 11.93 A ATOM 705 CE MET A 102 47.530 4.281 22.436 1.0010.87 A ATOM 706 C MET A 102 44.645 5.163 17.563 1.00 19.65 A ATOM 707 OMET A 102 43.829 4.371 17.091 1.00 19.47 A ATOM 708 N ILE A 103 44.3846.451 17.755 1.00 18.90 A ATOM 709 CA ILE A 103 43.108 7.049 17.406 1.0021.34 A ATOM 710 CB ILE A 103 42.959 8.459 18.055 1.00 20.40 A ATOM 711CG2 ILE A 103 41.789 9.217 17.425 1.00 19.03 A ATOM 712 CG1 ILE A 10342.757 8.312 19.565 1.00 19.84 A ATOM 713 CD1 ILE A 103 42.941 9.60120.345 1.00 18.82 A ATOM 714 C ILE A 103 43.045 7.183 15.881 1.00 24.13A ATOM 715 O ILE A 103 41.996 6.968 15.271 1.00 24.18 A ATOM 716 N TYR A104 44.182 7.530 15.277 1.00 27.06 A ATOM 717 CA TYR A 104 44.283 7.70113.827 1.00 29.10 A ATOM 718 CB TYR A 104 45.702 8.155 13.462 1.00 30.33A ATOM 719 CG TYR A 104 45.914 9.635 13.689 1.00 30.58 A ATOM 720 CD1TYR A 104 47.148 10.137 14.093 1.00 29.56 A ATOM 721 CE1 TYR A 10447.328 11.499 14.323 1.00 29.30 A ATOM 722 CD2 TYR A 104 44.862 10.53313.516 1.00 31.51 A ATOM 723 CE2 TYR A 104 45.032 11.892 13.742 1.0031.90 A ATOM 724 CZ TYR A 104 46.264 12.366 14.146 1.00 30.22 A ATOM 725OH TYR A 104 46.411 13.711 14.379 1.00 32.03 A ATOM 726 C TYR A 10443.895 6.466 13.017 1.00 29.49 A ATOM 727 O TYR A 104 43.336 6.58711.926 1.00 30.07 A ATOM 728 N ARG A 105 44.188 5.287 13.553 1.00 30.44A ATOM 729 CA ARG A 105 43.851 4.039 12.883 1.00 31.09 A ATOM 730 CB ARGA 105 44.714 2.889 13.407 1.00 32.59 A ATOM 731 CG ARG A 105 46.0982.802 12.783 1.00 35.48 A ATOM 732 CD ARG A 105 46.758 1.479 13.140 1.0038.68 A ATOM 733 NE ARG A 105 47.041 1.380 14.572 1.00 41.14 A ATOM 734CZ ARG A 105 48.198 1.727 15.132 1.00 42.11 A ATOM 735 NH1 ARG A 10549.192 2.196 14.382 1.00 41.27 A ATOM 736 NH2 ARG A 105 48.362 1.60516.446 1.00 42.61 A ATOM 737 C ARG A 105 42.382 3.705 13.104 1.00 31.15A ATOM 738 O ARG A 105 41.887 2.692 12.599 1.00 30.58 A ATOM 739 N ASN A106 41.691 4.552 13.867 1.00 30.35 A ATOM 740 CA ASN A 106 40.272 4.34914.137 1.00 31.38 A ATOM 741 CB ASN A 106 39.995 4.297 15.639 1.00 30.13A ATOM 742 CG ASN A 106 40.393 2.984 16.250 1.00 30.74 A ATOM 743 OD1ASN A 106 41.568 2.754 16.549 1.00 30.89 A ATOM 744 ND2 ASN A 106 39.4152.094 16.427 1.00 30.12 A ATOM 745 C ASN A 106 39.408 5.429 13.517 1.0032.04 A ATOM 746 O ASN A 106 38.293 5.677 13.970 1.00 30.99 A ATOM 747 NLEU A 107 39.914 6.070 12.474 1.00 34.74 A ATOM 748 CA LEU A 107 39.1447.115 11.828 1.00 39.45 A ATOM 749 CB LEU A 107 39.166 8.383 12.694 1.0039.46 A ATOM 750 CG LEU A 107 40.507 8.908 13.220 1.00 38.61 A ATOM 751CD1 LEU A 107 41.418 9.298 12.068 1.00 38.52 A ATOM 752 CD2 LEU A 10740.252 10.105 14.120 1.00 37.16 A ATOM 753 C LEU A 107 39.614 7.44110.425 1.00 42.25 A ATOM 754 O LEU A 107 40.442 6.732 9.848 1.00 42.75 AATOM 755 N VAL A 108 39.057 8.516 9.882 1.00 46.00 A ATOM 756 CA VAL A108 39.397 8.997 8.549 1.00 49.43 A ATOM 757 CB VAL A 108 38.365 8.5417.499 1.00 48.08 A ATOM 758 CG1 VAL A 108 39.032 8.443 6.146 1.00 48.02A ATOM 759 CG2 VAL A 108 37.735 7.216 7.907 1.00 46.78 A ATOM 760 C VALA 108 39.389 10.529 8.612 1.00 53.19 A ATOM 761 O VAL A 108 38.32111.145 8.684 1.00 52.78 A ATOM 762 N VAL A 109 40.576 11.136 8.601 1.0057.34 A ATOM 763 CA VAL A 109 40.708 12.596 8.664 1.00 61.10 A ATOM 764CB VAL A 109 42.177 13.052 8.480 1.00 61.46 A ATOM 765 CG1 VAL A 10942.306 14.528 8.847 1.00 61.63 A ATOM 766 CG2 VAL A 109 43.115 12.1939.320 1.00 62.28 A ATOM 767 C VAL A 109 39.875 13.293 7.584 1.00 63.47 AATOM 768 O VAL A 109 40.196 13.206 6.394 1.00 63.73 A ATOM 769 N VAL A110 38.817 13.989 8.004 1.00 65.46 A ATOM 770 CA VAL A 110 37.941 14.6977.073 1.00 67.21 A ATOM 771 CB VAL A 110 36.927 15.603 7.840 1.00 66.97A ATOM 772 CG1 VAL A 110 35.873 16.155 6.878 1.00 67.04 A ATOM 773 CG2VAL A 110 36.262 14.817 8.961 1.00 66.24 A ATOM 774 C VAL A 110 38.77515.570 6.119 1.00 69.20 A ATOM 775 O VAL A 110 39.949 15.862 6.458 1.0069.94 A ATOM 776 OXT VAL A 110 38.248 15.956 5.046 1.00 70.58 A ATOM 777C1 CID A 1 46.320 13.011 27.769 1.00 21.13 INH1 ATOM 778 C2 CID A 146.849 12.548 26.529 1.00 21.31 INH1 ATOM 779 C3 CID A 1 46.319 13.05325.307 1.00 21.72 INH1 ATOM 780 C4 CID A 1 45.283 14.011 25.313 1.0021.74 INH1 ATOM 781 C5 CID A 1 44.753 14.472 26.542 1.00 21.42 INH1 ATOM782 C6 CID A 1 45.261 13.981 27.804 1.00 22.49 INH1 ATOM 783 C7 CID A 144.698 14.428 29.194 1.00 23.66 INH1 ATOM 784 C8 CID A 1 44.052 15.86029.277 1.00 25.69 INH1 ATOM 785 O1 CID A 1 43.517 16.308 30.288 1.0029.01 INH1 ATOM 786 O2 CID A 1 44.409 16.731 28.309 1.00 30.64 INH1 ATOM787 N1 CID A 1 43.714 13.347 29.673 1.00 19.99 INH1 ATOM 788 C9 CID A 142.570 12.996 28.781 1.00 18.95 INH1 ATOM 789 C10 CID A 1 42.648 11.64128.034 1.00 18.04 INH1 ATOM 790 C11 CID A 1 43.669 10.654 28.259 1.0017.01 INH1 ATOM 791 C12 CID A 1 43.700 9.448 27.530 1.00 16.97 INH1 ATOM792 C13 CID A 1 42.708 9.210 26.564 1.00 17.64 INH1 ATOM 793 CL1 CID A 142.737 7.786 25.674 1.00 14.96 INH1 ATOM 794 C14 CID A 1 41.687 10.15326.318 1.00 18.96 INH1 ATOM 795 C15 CID A 1 41.659 11.354 27.046 1.0017.55 INH1 ATOM 796 C16 CID A 1 41.205 13.214 29.506 1.00 18.08 INH1ATOM 797 O3 CID A 1 40.363 13.973 29.015 1.00 18.82 INH1 ATOM 798 N2 CIDA 1 40.909 12.577 30.698 1.00 16.78 INH1 ATOM 799 C17 CID A 1 41.69011.680 31.447 1.00 17.03 INH1 ATOM 800 C18 CID A 1 43.101 11.735 31.5721.00 16.70 INH1 ATOM 801 C19 CID A 1 43.977 12.786 30.947 1.00 19.45INH1 ATOM 802 O4 CID A 1 44.965 13.160 31.626 1.00 18.52 INH1 ATOM 803C20 CID A 1 43.769 10.744 32.353 1.00 17.26 INH1 ATOM 804 C21 CID A 143.051 9.726 33.006 1.00 18.91 INH1 ATOM 805 I1 CID A 1 44.145 8.34334.119 1.00 19.64 INH1 ATOM 806 C22 CID A 1 41.657 9.670 32.902 1.0017.86 INH1 ATOM 807 C23 CID A 1 40.966 10.637 32.123 1.00 17.23 INH1ATOM 808 CL2 CID A 1 46.930 12.505 23.805 1.00 20.99 INH1 ATOM 809 CL3CID A 1 40.751 8.456 33.699 1.00 20.31 INH1 END

We claim:
 1. A crystal comprising HDM2, or a fragment, or targetstructural motif or derivative thereof, and a ligand, wherein saidligand is a small molecule inhibitor.
 2. The crystal of claim 1 whereinsaid fragment or derivative thereof is a peptide selected from the groupconsisting of SEQ ID NO: 1 (amino acid sequence of full length HDM2),SEQ ID NO: 2 (amino acid residues 17-111 of SEQ ID NO: 1), SEQ ID NO. 3(amino acid residues 23-114 of SEQ ID NO: 1) and SEQ ID NO. 4 (Gly¹⁶-SEQID NO: 2).
 3. The crystal of claim 1 wherein said crystal has aspacegroup selected from the group consisting of a trigonal spacegroupof P3₂21 and a tetragonal spacegroup of P4₃2₁2.
 4. The crystal of claim1, wherein the crystal effectively diffracts X-rays for determination ofatomic coordinates to a resolution of at least about 3.0 Å.
 5. Thecrystal of claim 1, wherein the ligand is in crystalline form.
 6. Thecrystal of claim 1 wherein said ligand is selected from the groupconsisting of(4-Chloro-phenyl)-[3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-aceticacid;[8-Chloro-3-(4-chloro-phenyl)-7-iodo-2,5-dioxo-1,2,3,5-tetrahydro-benzo[e][1,4]diazepin-4-yl]-(4-chloro-phenyl)-aceticacid); and derivatives thereof.
 7. The crystal of claim 1 wherein saidHDM2 comprises a peptide having at least 95% sequence identity to SEQ IDNO.
 2. 8. A crystal comprising SEQ ID NO: 2 comprising an atomicstructure characterized by the coordinates of Table 1 or Table
 2. 9. Thecrystal of claim 1 comprising a unit cell having dimensions selectedfrom the group consisting of: dimensions of about 98.6 Å, 98.6 Å and74.7 Å, and about alpha=90°, beta=90° and gamma=120°; and, dimensions ofabout 54.3 Å, 54.3 Å, 83.3 Å and about alpha=90°, beta=90° andgamma=90°.
 10. A computer system comprising: (a) a database containinginformation on the three dimensional structure of a crystal comprisingHDM2, or a fragment or a target structural motif or derivative thereof,and a ligand, wherein said ligand is a small molecule inhibitor, storedon a computer readable storage medium; and, (b) a user interface to viewthe information.
 11. A computer system of claim 10, wherein theinformation comprises diffraction data obtained from a crystalcomprising SEQ ID NO:2.
 12. A computer system of claim 10, wherein theinformation comprises an electron density map of a crystal formcomprising SEQ ID NO:2.
 13. A computer system of claim 10, wherein theinformation comprises the structure coordinates of Table 1 or Table 2 orhomologous structure coordinates comprising a root mean square deviationof non-hydrogen atoms of less than about 1.5 Å when superimposed on thenon-hydrogen atom positions of the corresponding atomic coordinates ofTable 1 or Table
 2. 14. A computer system of claim 13, wherein theinformation comprises structure coordinates for amino acid residuescomprising a root mean square deviation of non-hydrogen atoms of lessthan about 0.75 Å when superimposed on the non-hydrogen atom positionsof the corresponding atomic coordinates of Table 1 or Table
 2. 15. Acomputer system of claim 10, wherein the information comprises thestructure coordinates for amino acids Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷according to Table 1 or Table 2 or similar structure coordinates forsaid amino acids comprising a root mean square deviation of non-hydrogenatoms of less than about 1.5 Å when superimposed on the non-hydrogenatom positions of the corresponding atomic coordinates of Table 1 orTable
 2. 16. A computer system of claim 15, wherein the informationfurther comprises the structure coordinates for amino acids Val⁵³,Leu⁵⁴, Phe⁵⁵, Leu⁵⁷, Gly⁵⁸, Gln⁵⁹, Ile⁶², Met⁶², Tyr⁶⁷, Gln⁷², His⁷³,Ile⁷⁴, Val⁷⁵, Phe⁸⁶, Phe⁹¹, Val⁹³, Lys⁹⁴, Glu⁹⁵, His⁹⁶, Ile⁹⁹, Tyr¹⁰⁰,Ile¹⁰³ according to Table 1 or or Table 2 or similar structurecoordinates for said amino acids comprising a root mean square deviationof non-hydrogen atoms of less than about 1.5 Å when superimposed on thenon-hydrogen atom positions of the corresponding atomic coordinates ofTable 1 or Table
 2. 17. A method of evaluating the potential of an agentto associate with HDM2 comprising: (a) exposing HDM2 to the agent; and(b) detecting the association of said agent to HDM2 amino acid residuesSer¹⁷, Ile¹⁹, Leu82 and Arg⁹⁷ thereby evaluating the potential.
 18. Amethod of claim 17, wherein the agent is a virtual compound.
 19. Amethod of evaluating the potential of an agent to associate with thepeptide having aa¹⁶ -SEQ ID NO: 2, comprising: (a) exposing aa¹⁶-SEQ IDNO: 2 to the agent; and (b) detecting the level of association of theagent to aa¹⁶-SEQ ID NO: 2, thereby evaluating the potential.
 20. Amethod of claim 19, wherein the agent is a virtual compound.
 21. Amethod of claim 17 wherein step (a) comprises comparing the atomicstructure of the compound to the three dimensional structure of HDM2.22. A method of claim 17, wherein the comparing comprises employing acomputational means to perform a fitting operation between the compoundand at least one binding site of HDM2.
 23. A method of claim 22, whereinthe binding site is defined by structure coordinates for amino acidsSer¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷ according to Table 1 or Table 2 or similarstructure coordinates for said amino acids comprising a root mean squaredeviation of non-hydrogen atoms of less than about 1.5 Å whensuperimposed on the non-hydrogen atom positions of the correspondingatomic coordinates of Table 1 or Table
 2. 24. A method of claim 23,wherein the binding site is further defined by structure coordinates foramino acids Val⁵³, Leu⁵⁴, Phe⁵⁵, Leu⁵⁷, Gly⁵⁸, Gln⁵⁹, Ile⁶¹, Met⁶²,Tyr⁶⁷, Gln⁷², His⁷³, Ile⁷⁴, Val⁷⁵, Phe⁸⁶, Phe⁹¹, Val⁹³, Lys⁹⁴, Glu⁹⁵,His⁹⁶, Ile⁹⁹, Tyr¹⁰⁰, Ile¹⁰³ according to Table 1 or Table 2 or similarstructure coordinates for said amino acids comprising a root mean squaredeviation of non-hydrogen atoms of less than about 1.5 Å whensuperimposed on the non-hydrogen atom positions of the correspondingatomic coordinates of Table 1 or Table
 2. 25. A method of claim 17,wherein the agent is exposed to crystalline SEQ ID NO:2 and thedetecting of step (b) comprises determining the three dimensionalstructure of the agent-SEQ ID NO: 2 complex.
 26. A method of identifyinga potential agonist or antagonist against HDM2 comprising: (a) employingthe three dimensional structure of HDM2 cocrystallized with a smallmolecule inhibitor to design or select said potential agonist orantagonist.
 27. A method of claim 26, wherein the three dimensionalstructure corresponds to the atomic structure characterized by thecoordinates of Table 1 or Table 2 or similar structure coordinatescomprising a root mean square deviation of non-hydrogen atoms of lessthan about 1.5 Å when superimposed on the non-hydrogen atom positions ofthe corresponding atomic coordinates of Table 1 or Table
 2. 28. A methodof claim 26, further comprising the steps of: (b) synthesizing thepotential agonist or antagonist; and (c) contacting the potentialagonist or antagonist with HDM2.
 29. A method of locating the attachmentsite of an inhibitor to HDM2, comprising: (a) obtaining X-raydiffraction data for a crystal of HDM2; (b) obtaining X-ray diffractiondata for a complex of HDM2 and an inhibitor; (c) subtracting the X-raydiffraction data obtained in step (a) from the X-ray diffraction dataobtained in step (b) to obtain the difference in the X-ray diffractiondata; (d) obtaining phases that correspond to X-ray diffraction dataobtained in step (a); (e) utilizing the phases obtained in step (d) andthe difference in the X-ray diffraction data obtained in step (c) tocompute a difference Fourier image of the inhibitor; and, (f) locatingthe attachment site of the inhibitor to HDM2 based on the computationsobtained in step (e).
 30. A method of obtaining a modified inhibitorcomprising: (a) obtaining a crystal comprising HDM2 and an inhibitor;(b) obtaining the atomic coordinates of the crystal; (c) using theatomic coordinates and one or more molecular modeling techniques todetermine how to modify the interaction of the inhibitor with HDM2; and(d) modifying the inhibitor based on the determinations obtained in step(c) to produce a modified inhibitor.
 31. The method of claim 30 whereinsaid crystal comprises a peptide selected from the group consisting of:a peptide having SEQ ID NO: 2; a peptide having SEQ ID NO: 3 and apeptide having SEQ ID NO:4.
 32. A method of claim 30, wherein the one ormore molecular modeling techniques are selected from the groupconsisting of graphic molecular modeling and computational chemistry.33. A method of claim 30, wherein step (a) comprises detecting theinteraction of the inhibitor to HDM2 amino acid residues Ser¹⁷, Ile¹⁹,Leu⁸² and Arg⁹⁷.
 34. An HDM2 inhibitor identified by the method of claim30.
 35. An isolated protein fragment comprising a binding pocket oractive site defined by structure coordinates of HDM2 amino acid residuesSer¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷.
 36. An isolated fragment of claim 35linked to a solid support.
 37. An isolated nucleic acid moleculeencoding the fragment of claim
 35. 38. A vector comprising a nucleicacid molecule of claim
 37. 39. A host cell comprising the vector ofclaim
 38. 40. A method of producing a protein fragment, comprisingculturing the host cell of claim 39 under conditions in which thefragment is expressed.
 41. A method of screening for an agent thatassociates with HDM2, comprising: (a) exposing a protein moleculefragment of claim 35 to the agent; and (b) detecting the level ofassociation of the agent to the fragment.
 42. A kit comprising a proteinmolecule fragment of claim
 35. 43. A method for the production of acrystal complex comprising an HDM2 polypeptide-ligand comprising: (a)contacting the HDM2 polypeptide with said ligand in a suitable solutioncomprising PEG and NaSCN; and, b) crystallizing said resulting complexof HDM2 polypeptide-ligand from said solution.
 44. The method of claim43 wherein said HDM2 polypeptide is a polypeptide having SEQ ID NO: 2.45. The method of claim 43 wherein said PEG has an average molecularweight range from 100 to 1000, wherein said PEG is present in solutionat a range from about 0.5% w/v to about 10% w/v and said NaSCN ispresent in solution at a range of from about 50 mM to about 150 mM. 46.The method of claim 45 wherein said PEG has an average molecular weightof about 400 and is present in solution at about 2% w/v and said NaSCNis present in solution at about 100 mM.
 47. The method of claim 46wherein said solution further comprises about 1.8-2.4 M (NH₄)₂SO₄ andabout 100 mM buffer.
 48. A method for the production of a crystal ofclaim 1 comprising crystallizing a peptide comprising a sequenceselected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ IDNO: 3 and SEQ ID NO:4 with a potential inhibitor.
 49. A method foridentifying a potential inhibitor of HDM2 comprising: a) using a threedimensional structure of HDM2 as defined by atomic coordinates accordingto Table 1 or Table 2; b) replacing one or more HDM2 amino acidsselected from Ser¹⁷, Ile¹⁹, Leu⁸² and Arg⁹⁷ in said three-dimensionalstructure with a different amino acid to produce a modified HDM2; c)using said three-dimensional structure to design or select saidpotential inhibitor; d) synthesizing said potential inhibitor; and, e)contacting said potential inhibitor with said modified HDM2 in thepresence of a substrate to test the ability of said potential inhibitorto inhibit HDM2 or said modified HDM2.
 50. The method of claim 49wherein said replacing one or more amino acid residues further comprisesreplacing SEQ ID NO: 2 amino acids selected from the group consisting ofVal⁵³, Leu⁵⁴, Phe⁵⁵, Leu⁵⁷, Gly⁵⁸, Gln⁵⁹, Ile⁶¹, Met⁶², Tyr⁶⁷, Gln⁷²,His⁷³, Ile⁷⁴, Val⁷⁵, Phe⁸⁶, Phe⁹¹, Val⁹³, Lys⁹⁴, Glu⁹⁵, His⁹⁶, Ile⁹⁹,Tyr¹⁰⁰, and Ile¹⁰³.
 51. The method of claim 49 wherein said potentialinhibitor is selected from a database.
 52. The method of claim 49wherein said potential inhibitor is designed de novo.
 53. The method ofclaim 49 wherein said potential inhibitor is designed from a knowninhibitor.
 54. The method of claim 49, wherein said step of employingsaid three-dimensional structure to design or select said potentialinhibitor comprises the steps of: a) identifying chemical entities orfragments capable of associating with modified HDM2; and b) assemblingthe identified chemical entities or fragments into a single molecule toprovide the structure of said potential inhibitor.
 55. The method ofclaim 49, wherein the potential inhibitor is a competitive inhibitor ofSEQ ID NO:4 (Gly¹⁶-SEQ ID NO: 2).
 56. The method of claim 49, whereinsaid potential inhibitor is a non-competitive or uncompetitive inhibitorof SEQ ID NO:4 (Gly¹⁶-SEQ ID NO: 2).
 57. The inhibitor identified by themethod of claim 49.